Methods, Systems and Computer Program Products for Non-Invasive Determination of Blood Flow Distribution Using Speckle Imaging Techniques and Hemodynamic Modeling

ABSTRACT

Non-invasive methods for determining blood flow distribution in a region of interest are provided. The method includes illuminating a region of interest of a subject with a coherent light source; sequentially acquiring at least two speckle images of the region of interest, wherein sequentially acquiring the at least two speckle images comprises acquiring the at least two speckle images in synchronization with motion of the heart of the subject; and electronically processing the at least two acquired speckle images based on the temporal variation of the pixel intensities in the at least two acquired speckle images to generate a laser speckle contrast imaging (LSCI) image, determine distribution of blood flow speed in principal vessels and quantify perfusion distribution in tissue in the region of interest from the LSCI image. The LSCI image enables detection of different blood flow speeds.

CLAIM OF PRIORITY

The present application claims priority from U.S. application Ser. No.13/819,817, filed Feb. 28, 2013, which is a U.S. national phaseapplication of PCT/US2012/020626, filed Jan. 9, 2012, which claimspriority to U.S. Provisional Application No. 61/431,161, filed Jan. 10,2011 and U.S. Provisional Application No. 61/476,854, filed Apr. 19,2011, the disclosures of which are hereby incorporated herein byreference as if set forth in their entirety.

RESERVATION OF COPYRIGHT

A portion of the disclosure of this patent document contains materialwhich is subject to copyright protection. The copyright owner, EastCarolina University of Greenville, N.C., has no objection to thereproduction by anyone of the patent document or the patent disclosure,as it appears in the Patent and Trademark Office patent file or records,but otherwise reserves all copyright rights whatsoever.

FIELD

The present inventive concept relates generally to determination ofblood flow distribution and, more particularly, to the use of speckleimaging techniques for non-invasive determination of blood flowdistribution.

BACKGROUND

Revascularization is an interventional procedure for the provision of anew, additional, or augmented blood supply to a body part or organ.Revascularization typically involves a thorough analysis and/ordiagnosis and treatment of the existing diseased vasculature of theaffected organ. In some instances, revascularization can be aided by theuse of different imaging modalities, such as magnetic resonance imaging(MRI), positron emission tomography (PET) scan, computed tomography (CT)scan, and X ray fluoroscopy.

Revascularization is designed to improve blood flow to tissues perfusedby the principal arterial vessel(s) supplying that tissue.Revascularization may be needed, for example, due to an obstruction inthe native arterial vessel supplying that tissue. Coronary artery bypassgrafting (CABG) is a revascularization procedure that may be used toincrease blood flow to ischemic myocardium by bypassing the nativecoronary obstructions.

There are two measurement components to the revascularizationevaluation, blood flow in the principal arterial supply and quantitativeperfusion in the tissue. Conventional methods for measurement of bloodflow and perfusion are limited, despite the benefit these measurementswould bring to the clinical evaluation of the quality of therevascularization procedure.

Some conventional interoperative methods of measuring blood flow arebased on ultrasound detection of blood flow in the graft conduits, butnot the native principal arterial vessel(s). Some conventionalangiographic evaluation methods include conventional coronaryangiography performed in a hybrid operating room setting at the time ofsurgery. Recently, Novadaq Technologies, Inc. of Toronto, Canada hasintroduced fluorescence imaging that uses both angiographic imageevaluation and quantitative perfusion evaluation to CABG.

However, ultrasound detection typically requires physical contactbetween the graft vessel and a probe. Furthermore, ultrasound detectiontypically relies on proper placement of the probe around the vessel toobtain accurate measurement of flow speed and can be unreliable,measurement to measurement.

Coronary angiography typically requires radiation and administration oftoxic image contrast agent. Furthermore, hybrid operating rooms used forcoronary angiography can be relatively expensive, making this methodunavailable to many patients undergoing CABG.

Fluorescence imaging typically requires injection of non-toxic dye intothe patient. Furthermore, fluorescence imaging typically cannot provideinformation to directly determine the speed of blood flow in principalvessels. Despite the above, there remains a need for alternative methodsof determining blood flow.

SUMMARY

Some embodiments of the present inventive concept provide a non-invasivemethod for measuring blood flow in principal vessels of a heart of asubject, the method including illuminating a region of interest in theheart with a coherent light source, wherein the coherent light sourcehas a wavelength of from about 600 nm to about 1100 nm; sequentiallyacquiring at least two speckle images of the region of interest in theheart during a fixed time period, wherein sequentially acquiring the atleast two speckle images comprises acquiring the at least two speckleimages in synchronization with motion of the heart of the subject; andelectronically processing the at least two acquired speckle images basedon the temporal variation of the pixel intensities in the at least twoacquired speckle images to generate a laser speckle contrast imaging(LSCI) image and determine spatial distribution of blood flow speed inthe principal vessels and perfusion distribution in tissue in the regionof interest in the heart from the LSCI image.

In further embodiments, sequentially acquiring the at least two speckleimages may include electronically monitoring an EKG cardiac cycle of thesubject; and electronically synchronizing acquisition of speckle imageswith the EKG signals.

In still further embodiments, the sequentially acquiring and theelectronically evaluating may be performed before a procedure performedon a subject and after the procedure performed on the subject. Themethod may further include comparing the determined blood flow speed inthe principal vessels and perfusion distribution in tissue in the regionof interest in the heart before the procedure with the determined bloodflow speed in the principal vessels and perfusion distribution in tissuein the region of interest in the heart after the procedure to accesssuccess of the procedure.

In some embodiments, the method further includes calculating a velocityfield for the region of interest in the heart; calculating blood flowspeed in the region of interest in the heart based on the calculatedvelocity field; and comparing the calculated blood flows speed in theregion of interest to the blood flow speed determined using the acquiredat least two speckle images of the region of interest in the heart toverify results obtained using the at least two speckle images.

In further embodiments, the velocity field is calculated using equations(9) and (10) set out below.

In still further embodiments, the coherent light source may have awavelength of from about 600 nm to about 1100 nm and may allowrelatively deep penetration of light into tissues to thereby allow anaccurate determination of blood flow speed in the principal vessels andthe perfusion distribution.

In some embodiments, the coherent light source may include a laserconfigured to illuminate the region of interest with a substantiallyconstant intensity. The laser may have a fixed or variable wavelength offrom about 600 nm to about 1100 nm. The laser may generate a laser beamhaving a substantially constant intensity within a field-of-view (FOV)of an imaging unit. The laser may be a low power and continuous-wavelaser such that the subject does not require any protective apparatus toshield the subject from effects of the laser.

In further embodiments, data acquisition may include sequentiallyacquiring from about 50 to about 1000 speckle images using the cameraduring the fixed time period of from about 1 ms to about 200 ms.

In still further embodiments, sequentially acquiring may includeacquiring from about 200 to about 500 speckle images during the fixedtime period.

In some embodiments, the fixed time period may be selected based on insitu acquisition of blood flow speed of the subject in the region ofinterest.

Further embodiments of the present inventive concept provide anon-invasive method for measuring blood flow in principal vessels of asubject, the method including illuminating a region of interest with acoherent light source, wherein the coherent light source has awavelength of from about 600 nm to about 1100 nm; sequentially acquiringat least two speckle images of the region of interest during a fixedtime period; electronically processing the at least two acquired speckleimages based on the temporal variation of the pixel intensities in theat least two acquired speckle images to generate a laser specklecontrast imaging (LSCI) image, and determine spatial distribution ofblood flow speed in the principal vessels and quantify perfusiondistribution in tissue in the region of interest from the LSCI image;calculating a velocity field for the region of interest; calculatingblood flow rate in the region of interest based on the calculatedvelocity field; and comparing the calculated blood flow speed in theregion of interest to the blood flow speed determined using the acquiredat least two speckle images of the region of interest to verify resultsobtained using the at least two speckle images.

Still further embodiments of the present inventive concept provide anon-invasive system for measuring blood flow in principal vessels in aheart of a subject, the system including a coherent light sourceconfigured to illuminate a region of interest in the heart of thesubject, the coherent light source having a wavelength of from about 600nm to about 1100 nm. A camera in communication with the coherent lightsource is provided that is configured to sequentially acquire at leasttwo speckle images of the region of interest in the heart during a fixedtime period, wherein acquisition of the at least two speckle images issynchronized with motion of the heart of the subject. A data processingcircuit is also provided that is configured to evaluate the temporalvariation of the pixel intensities in the at least two acquired speckleimages to generate an LSCI image and determine spatial distribution ofblood flow speed in the principal vessels and quantify perfusiondistribution in tissue in the region of interest in the heart from theLSCI image.

Some embodiments of the present inventive concept provide a computerprogram product for measuring blood flow in principal vessels in a heartof a subject, the computer program product comprising a non-transitorycomputer-readable storage medium having computer-readable program codeembodied in the medium. The computer-readable program code includescomputer readable program code configured to electronically evaluatetemporal variation of the pixel intensities in the at least two acquiredspeckle images to generate an LSCI image and determine spatialdistribution of blood flow speed in the principal vessels and quantifyperfusion distribution in tissue in the region of interest in the heartfrom the LSCI image, wherein the at least two speckle images aresequentially acquired using a camera during a fixed time period when theregion of interest of the subject is illuminated by a coherent lightsource having a wavelength of from about 600 nm to about 1100 nm; andcomputer readable program code configured to sequentially acquire the atleast two speckle images in synchronization with motion of the heart ofthe subject.

Further embodiments of the present inventive concept provide anon-invasive method for determining blood flow distribution in a regionof interest. The method includes illuminating a region of interest of asubject with a coherent light source; sequentially acquiring at leasttwo speckle images of the region of interest, wherein sequentiallyacquiring the at least two speckle images comprises acquiring the atleast two speckle images in synchronization with motion of the heart ofthe subject; and electronically processing the at least two acquiredspeckle images based on the temporal variation of the pixel intensitiesin the at least two acquired speckle images to generate a laser specklecontrast imaging (LSCI) image, determine distribution of blood flowspeed in principal vessels and quantify perfusion distribution in tissuein the region of interest from the LSCI image. The LSCI image enablesdetection of different blood flow speeds.

In still further embodiments, acquiring the at least two speckle imagesin synchronization with motion of the heart of the subject may furtherinclude electronically monitoring an EKG cardiac cycle of the subject;and electronically synchronizing acquisition of speckle images with theEKG signals.

In some embodiments, the region of interest may be a beating heart andthe method may further include generating instantaneous flow and/orperfusion maps for the region of interest at any time during a cardiaccycle using the EKG signals to synchronize data acquisition. Theinstantaneous flow and/or perfusion maps generated at first and secondtimes may be compared to determine efficacy of a treatment. In certainembodiments, the first time may be a time before a treatment isadministered and the second time may be a time after the treatment isadministered.

In further embodiments, the region of interest may be a beating heartand the method may further include generating average flow and/orperfusion maps for the region of interest over two or more cardiaccycles using EKG signals to synchronize data acquisition. Average flowand/or perfusion maps generated at first and second times may becompared to determine efficacy of a treatment. In certain embodiments,the first time may be a time before a treatment is administered and thesecond time may be a time after the treatment is administered.

In still further embodiments, the region of interest may be anon-cardiac region of the subject and the method may further includegenerating instantaneous flow and/or perfusion maps at any time duringdata acquisition using the EKG signals to synchronize data acquisition.Instantaneous flow and/or perfusion maps generated at first and secondtimes may be compared to determine efficacy of a treatment. The firsttime may be a time before a treatment is administered and the secondtime may be a time after the treatment is administered.

In some embodiments, the region of interest may be a non-cardiac regionof the subject and the method may further include generating averageflow and/or perfusion maps over two or more periods of data acquisitionusing EKG signals to synchronize data acquisition. Average flow and/orperfusion maps generated at first and second times may be compared todetermine efficacy of a treatment. The first time may be a time before atreatment is administered and the second time may be a time after thetreatment is administered.

In further embodiments, the coherent light source may have a wavelengthof from about 600 nm to about 1100 nm.

Still further embodiments of the present inventive concept providenon-invasive methods for determining blood flow distribution in a regionof interest. The method includes illuminating a region of interest of asubject with a coherent light source; sequentially acquiring at leasttwo speckle images of the region of interest, wherein sequentiallyacquiring the at least two speckle images comprises acquiring the atleast two speckle images in synchronization with motion of the heart ofthe subject; electronically processing the at least two acquired speckleimages based on the temporal variation of the pixel intensities in theat least two acquired speckle images to generate a laser specklecontrast imaging (LSCI) image, determine distribution of blood flowspeed in principal vessels and quantify perfusion distribution in tissuein the region of interest from the LSCI image; generating one ofinstantaneous flow and/or perfusion maps for the region of interest atany time during data acquisition using EKG signals to synchronize dataacquisition and average flow and/or perfusion maps over two or moreperiods of data acquisition using the EKG signals to synchronize dataacquisition; and comparing one of the instantaneous flow and/orperfusion maps and the average flow and/or perfusion maps generated atfirst and second times to determine efficacy of a treatment.

Some embodiments of the present inventive concept provide non-invasivemethods for determining blood flow distribution in a region of interest.The method may include illuminating a region of interest of a subjectwith a coherent light source; sequentially acquiring at least twospeckle images of the region of interest, wherein sequentially acquiringthe at least two speckle images comprises acquiring the at least twospeckle images in synchronization in synchronization with anelectrocardiogram (EKG); selecting at least two speckle images usingEKG-based timing; processing the selected images to determine aninstantaneous/average flow speed (centimeter/second) in the region ofinterest using at least one of a temporal contrast algorithm and aspatial contrast algorithm, wherein the EKG is used at any time duringdata acquisition and analysis to select frames to be processed, tolocate the instantaneous flow speed in one or more EKG cycles, and/or totarget the beginning and ending time of average flow speed analyses;inputting the instantaneous/average flow speed image into the analysismodel to generate a flow rate (cubic centimeter/second) map in theprincipal vessels and a perfusion map in the microvascular structure;and generating a direction of flow and pressure.

Related systems and computer program products may also be provided.

It is noted that aspects of the inventive concept described with respectto some embodiments, may be incorporated in different embodimentsalthough not specifically described relative thereto. That is, allembodiments and/or features of any embodiment can be combined in any wayand/or combination. Applicant reserves the right to change anyoriginally filed claim and/or file any new claim accordingly, includingthe right to be able to amend any originally filed claim to depend fromand/or incorporate any feature of any other claim or claims although notoriginally claimed in that manner. These and other objects and/oraspects of the present inventive concept are explained in detail in thespecification set forth below. Further features, advantages and detailsof the present inventive concept will be appreciated by those ofordinary skill in the art from a reading of the figures and the detaileddescription of the embodiments that follow, such description beingmerely illustrative of the present inventive concept.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 is a block diagram of a non-invasive system for measuring bloodflow in principal vessels of a subject in accordance with someembodiments of the present inventive concept(s).

FIG. 2A is a block diagram of a data processing system according toembodiments of the present inventive concept(s).

FIG. 2B is a more detailed block diagram of the data processing systemillustrated in FIG. 2 in accordance with some embodiments of the presentinventive concept(s).

FIGS. 3 and 4 are flowcharts illustrating operations for measuring bloodflow in principal vessels in accordance with various embodiments of thepresent inventive concept(s).

FIG. 5 is a digital photograph of a system for measuring flow of bloodphantom used in an experiment performed in accordance with someembodiments of the present inventive concept(s).

FIGS. 6 through 9 are close up digital photographs of certain elementsin the system illustrated in FIG. 5 in accordance with some embodimentsof the present inventive concept(s).

FIGS. 10 and 11 are digital photographs of an exemplary flow generationsystem used in some embodiments of the present inventive concept(s).

FIG. 12 is a graph illustrating the change in height (cm) vs. flow rate(ml/min) in the flow generation system in accordance with someembodiments of the present inventive concept(s).

FIGS. 13A through 13D are digital images illustrating a “no flow” casein accordance with some embodiments of the present inventive concept(s).

FIGS. 14A through 14D are images illustrating a “flow 1” case inaccordance with some embodiments of the present inventive concept(s).

FIGS. 15A through 15D are images illustrating a “flow 2” case inaccordance with some embodiments of the present inventive concept(s).

FIGS. 16A through 16D are images illustrating a “flow 3” case inaccordance with some embodiments of the present inventive concept(s).

FIGS. 17A through 17D are images (averaged over a number of frames)illustrating a speckle image for each of four flow cases illustrated inFIGS. 13 through 16 in accordance with some embodiments of the presentinventive concept(s).

FIGS. 18A through 18D are inverted speckle contrast images illustratingeach of the four flow cases illustrated in FIGS. 13 through 16 inaccordance with some embodiments of the present inventive concept(s).

FIGS. 19A through 19D are graphs illustrating a vertical profile ofinverted speckle contrast images for each of the four flow cases inFIGS. 13 through 16 in accordance with some embodiments of the presentinventive concept(s).

FIG. 20 is a graph illustrating predicted flow rate (ml/min) vs.inverted speckle image pixel intensity in accordance with someembodiments of the present inventive concept(s).

FIG. 21 is a digital photograph of an exemplary system for measuringflow of blood phantom in accordance with some embodiments of the presentinventive concept(s).

FIGS. 22 through 27 are close up photographs of the elements in thesystem illustrated in FIG. 21 in accordance with some embodiments of thepresent inventive concept(s).

FIG. 28 is a graph illustrating flow rate (ml/min), flow speed (cm/min)and corresponding LAD flow rate vs. inverted speckle contrast imagepixel intensity in accordance with some embodiments of the presentinventive concept(s).

FIG. 29 is a graph illustrating flow speed vs. inverted speckle contrastimage pixel intensity in accordance with some embodiments of the presentinventive concept(s).

FIG. 30 is a graph illustrating average flow speed determined fromspeckle contrast images vs. flow speed determined from the pump flowrate in accordance with some embodiments of the present inventiveconcept(s).

FIGS. 31A, 31C and 31D are images illustrating effect of specularreflectance on acquired speckle image data in accordance with someembodiments of the present inventive concept(s).

FIG. 31B is a graph illustrating effect of specular reflectance onacquired speckle image data in accordance with some embodiments of thepresent inventive concept(s).

FIGS. 32A, 32C and 32D are images illustrating removal of specularreflectance of FIGS. 31A, 31C and 31D in accordance with someembodiments of the present inventive concept(s).

FIG. 32B is a graph illustrating removal of specular reflectance ofFIGS. 31A, 31B and 31D in accordance with some embodiments of thepresent inventive concept(s).

FIG. 33 is a digital photograph of an exemplary system for measuringblood flow including a reservoir used in an experiment performed inaccordance with some embodiments of the present inventive concept(s).

FIGS. 34A through 34D are digital images illustrating a “tube clamped”situation in accordance with some embodiments of the present inventiveconcept(s).

FIGS. 35A through 35D are images illustrating a “pump reading 100 mL”situation in accordance with some embodiments of the present inventiveconcept(s).

FIGS. 36A through 36D are images illustrating a “pump reading 500 mL”situation in accordance with some embodiments of the present inventiveconcept(s).

FIGS. 37A through 37D are images illustrating a “pump reading 1000 mL”situation in accordance with some embodiments of the present inventiveconcept(s).

FIGS. 38A through 38D are averaged speckle images (averaged over anumber of frames) for each of the four flow cases illustrated in FIGS.33 through 37 in accordance with some embodiments of the presentinventive concept(s).

FIGS. 39A through 39D are colorized inverted speckle contrast imagesillustrating each of the four flow case illustrated in FIGS. 33 through37 in accordance with some embodiments of the present inventiveconcept(s).

FIGS. 40A through 40D are graphs illustrating a vertical line profile ofthe inverted speckle contrast images for each of the four flow cases inFIGS. 33 through 37 in accordance with some embodiments of the presentinventive concept(s).

FIG. 41 is a graph illustrating flow speed (cm/min) vs. inverted specklecontrast image pixel intensity in accordance with some embodiments ofthe present inventive concept(s).

FIG. 42 is a graph illustrating flow speed (cm/min) vs. inverted specklecontrast image pixel intensity in accordance with some embodiments ofthe present inventive concept.

FIGS. 43A through 43D are images illustrating a “pump reading OmL”situation in accordance with some embodiments of the present inventiveconcept(s).

FIGS. 44A through 44D are images illustrating a “clamped pump” situationin accordance with some embodiments of the present inventive concept(s).

FIG. 45 is a diagram of a blood vessel having a narrowing in the middlethereof that can be accessed using methods and systems in accordancewith some embodiments of the present inventive concept.

FIG. 46 is a diagram illustrating velocity profiles of various locationsin the blood vessel illustrated in FIG. 45 obtained using flowhemodynamic modeling in accordance with some embodiments of the presentinventive concept.

FIG. 47 is a graph illustrating shear rate (which is related to flowrate) and horizontal coordinate (diameter) across the blood vesselillustrated in FIGS. 45 and 46 when the shear stress is about 1.0 secondin accordance with some embodiments of the present inventive concept.

FIG. 48 is a flowchart illustrating operations for measuring blood flowin principal vessels in accordance with various embodiments of thepresent inventive concept(s).

FIG. 49 is a diagram of a SPY device system used in accordance with someembodiments of the present inventive concept.

FIGS. 50A and 50B are images illustrating a single laser speckle imaging(LSI) image and a inverse laser speckle temporal contrast imaging(LSCTI) image, respectively, in accordance with some embodiments of thepresent inventive concept.

FIGS. 51A and 51B are images illustrating a single LSI image and ainverse LSCTI image, respectively, in accordance with some embodimentsof the present inventive concept.

FIGS. 52A and 52B are images illustrating a conventional SPY image and aLSCTI image, respectively, in accordance with some embodiments of thepresent inventive concept.

FIGS. 53A through 53D are images illustrating various aspects of the SPYsystem in accordance with some embodiments of the present inventiveconcept.

FIGS. 54A through 54C are images illustrating a single LSI image, alaser speckle spatial contrast imaging (LSSCI) image and an LSCTI image,respectively, of a non-beating human heart in accordance with variousembodiments of the present inventive concept.

FIGS. 55A through 55C are images illustrating a single LSI image, alaser speckle spatial contrast imaging (LSSCI) image and an LSCTI image,respectively, of a beating human heart in accordance with someembodiments of the present inventive concept.

FIG. 55D is a graph of average intensity vs. time illustrating a patternof movement of the beating heart in FIGS. 55A through 55C in accordancewith some embodiments of the present inventive concept.

FIG. 56A is a still image of a beating heart in accordance with someembodiments of the present inventive concept.

FIG. 56B is a graph of average intensity vs. time illustrating a patternof movement of the beating heart and using EKG gating in accordance withsome embodiments of the present inventive concept.

FIG. 57 is an example instantaneous flow and perfusion map that can begenerated at any time in a cardiac cycle using EKG synchronizationmethods in accordance with some embodiments of the present inventiveconcept.

FIGS. 58A through 58C are images illustrating a single LSI image, anLSSCI image and an LSCTI image, respectively, of a beating human heartat end diastole in accordance with some embodiments of the presentinventive concept.

FIG. 58D is a graph of average intensity vs. time illustrating aninstantaneous flow measurement of the beating heart in FIGS. 58A through58C in accordance with some embodiments of the present inventiveconcept.

FIGS. 59A through 59C are images illustrating a single LSI image, anLSSCI image and an LSCTI image, respectively, of a beating human heartat end systole in accordance with some embodiments of the presentinventive concept.

FIG. 59D is a graph of average intensity vs. time illustrating aninstantaneous flow measurement of the beating heart in FIGS. 59A through59C in accordance with some embodiments of the present inventiveconcept.

FIG. 60 is an example average flow and perfusion map that can begenerated in two or more cardiac cycles using EKG synchronizationmethods in accordance with some embodiments of the present inventiveconcept.

FIGS. 61A through 61C are images illustrating a single. LSI image, anLSSCI image and an LSCTI image, respectively, of a beating human heartat diastole in accordance with some embodiments of the present inventiveconcept.

FIG. 61D is a graph of average intensity vs. time illustrating anaverage flow measurement of the beating heart in FIGS. 61A through 61Cin accordance with some embodiments of the present inventive concept.

FIGS. 62A through 62C are images illustrating a single LSI image, anLSSCI image and an LSCTI image, respectively, of a beating human heartat systole in accordance with some embodiments of the present inventiveconcept.

FIG. 62D is a graph of average intensity vs. time illustrating anaverage flow measurement of the beating heart in FIGS. 62A through 62Cin accordance with some embodiments of the present inventive concept.

FIGS. 63A through 63C are images illustrating a single LSI image, anLSSCI image and an LSCTI image, respectively, of a beating human heartincluding a bypass graft that has been clamped in accordance with someembodiments of the present inventive concept.

FIG. 63D is a graph of average intensity vs. time of the beating heartin FIGS. 63A through 63C in accordance with some embodiments of thepresent inventive concept.

FIGS. 64A through 64C are images illustrating a single LSI image, anLSSCI image and an LSCTI image, respectively, of a beating human heartincluding a bypass graft where the clamp has been removed in accordancewith some embodiments of the present inventive concept.

FIG. 64D is a graph of average intensity vs. time of the beating heartin FIGS. 64A through 64C in accordance with some embodiments of thepresent inventive concept.

FIGS. 65A through 65C are images illustrating a single LSI image, anLSSCI image and an LSCTI image, respectively, of a human hand inaccordance with some embodiments of the present inventive concept.

FIGS. 66 and 67 are flowcharts illustrating operations in accordancewith various embodiments discussed herein.

DETAILED DESCRIPTION OF EMBODIMENTS

Embodiments of the present inventive concept will now be described morefully hereinafter with reference to the accompanying figures, in whichpreferred embodiments of the inventive concept are shown. This inventiveconcept may, however, be embodied in many different forms and should notbe construed as limited to the embodiments set forth herein. Likenumbers refer to like elements throughout. In the figures, layers,regions, elements or components may be exaggerated for clarity. Brokenlines illustrate optional features or operations unless specifiedotherwise.

The terminology used herein is for the purpose of describing particularembodiments only and is not intended to be limiting of the inventiveconcept. As used herein, the singular forms “a”, “an” and “the” areintended to include the plural forms as well, unless the context clearlyindicates otherwise. It will be further understood that the terms“comprises” and/or “comprising,” when used in this specification,specify the presence of stated features, integers, steps, operations,elements, and/or components, but do not preclude the presence oraddition of one or more other features, integers, steps, operations,elements, components, and/or groups thereof. As used herein, the term“and/or” includes any and all combinations of one or more of theassociated listed items. As used herein, phrases such as “between X andY” and “between about X and Y” should be interpreted to include X and Y.As used herein, phrases such as “between about X and Y” mean “betweenabout X and about Y.” As used herein, phrases such as “from about X toY” mean “from about X to about Y.”

Unless otherwise defined, all terms (including technical and scientificterms) used herein have the same meaning as commonly understood by oneof ordinary skill in the art to which this inventive concept belongs. Itwill be further understood that terms, such as those defined in commonlyused dictionaries, should be interpreted as having a meaning that isconsistent with their meaning in the context of the specification andrelevant art and should not be interpreted in an idealized or overlyformal sense unless expressly so defined herein. Well-known functions orconstructions may not be described in detail for brevity and/or clarity.

It will be understood that when an element is referred to as being “on”,“attached” to, “connected” to, “coupled” with, “contacting”, etc.,another element, it can be directly on, attached to, connected to,coupled with or contacting the other element or intervening elements mayalso be present. In contrast, when an element is referred to as being,for example, “directly on”, “directly attached” to, “directly connected”to, “directly coupled” with or “directly contacting” another element,there are no intervening elements present. It will also be appreciatedby those of skill in the art that references to a structure or featurethat is disposed “adjacent” another feature may have portions thatoverlap or underlie the adjacent feature.

It will be understood that, although the terms first, second, etc. maybe used herein to describe various elements, components, regions, layersand/or sections, these elements, components, regions, layers and/orsections should not be limited by these terms. These terms are only usedto distinguish one element, component, region, layer or section fromanother element, component, region, layer or section. Thus, a firstelement, component, region, layer or section discussed below could betermed a second element, component, region, layer or section withoutdeparting from the teachings of the inventive concept. The sequence ofoperations (or steps) is not limited to the order presented in theclaims or figures unless specifically indicated otherwise.

Spatially relative terms, such as “under”, “below”, “lower”, “over”,“upper” and the like, may be used herein for ease of description todescribe one element or feature's relationship to another element(s) orfeature(s) as illustrated in the figures. It will be understood that thespatially relative terms are intended to encompass differentorientations of the device in use or operation in addition to theorientation depicted in the figures. For example, if a device in thefigures is inverted, elements described as “under” or “beneath” otherelements or features would then be oriented “over” the other elements orfeatures. Thus, the exemplary term “under” can encompass both anorientation of over and under. The device may be otherwise oriented(rotated 90 degrees or at other orientations) and the spatially relativedescriptors used herein interpreted accordingly. Similarly, the terms“upwardly”, “downwardly”, “vertical”, “horizontal” and the like are usedherein for the purpose of explanation only unless specifically indicatedotherwise.

As will be appreciated by one of skill in the art, embodiments of thepresent inventive concept may be embodied as a method, system, dataprocessing system, or computer program product. Accordingly, the presentinventive concept may take the form of an embodiment combining softwareand hardware aspects, all generally referred to herein as a “circuit” or“module.” Furthermore, the present inventive concept may take the formof a computer program product on a non-transitory computer usablestorage medium having computer-usable program code embodied in themedium. Any suitable computer readable medium may be utilized includinghard disks, CD-ROMs, optical storage devices, or other electronicstorage devices.

Computer program code for carrying out operations of the presentinventive concept may be written in an object oriented programminglanguage such as Matlab, Mathematica, Java, Smalltalk, C or C++.However, the computer program code for carrying out operations of thepresent inventive concept may also be written in conventional proceduralprogramming languages, such as the “C” programming language or in avisually oriented programming environment, such as Visual Basic.

Certain of the program code may execute entirely on one or more of auser's computer, partly on the user's computer, as a stand-alonesoftware package, partly on the user's computer and partly on a remotecomputer or entirely on the remote computer. In the latter scenario, theremote computer may be connected to the user's computer through a localarea network (LAN) or a wide area network (WAN), or the connection maybe made to an external computer (for example, through the Internet usingan Internet Service Provider).

The inventive concept is described in part below with reference toflowchart illustrations and/or block diagrams of methods, devices,systems, computer program products and data and/or system architecturestructures according to embodiments of the inventive concept. It will beunderstood that each block of the illustrations, and/or combinations ofblocks, can be implemented by computer program instructions. Thesecomputer program instructions may be provided to a processor of ageneral-purpose computer, special purpose computer, or otherprogrammable data processing apparatus to produce a machine, such thatthe instructions, which execute via the processor of the computer orother programmable data processing apparatus, create means forimplementing the functions/acts specified in the block or blocks.

These computer program instructions may also be stored in acomputer-readable memory or storage that can direct a computer or otherprogrammable data processing apparatus to function in a particularmanner, such that the instructions stored in the computer-readablememory or storage produce an article of manufacture includinginstruction means which implement the function/act specified in theblock or blocks.

The computer program instructions may also be loaded onto a computer orother programmable data processing apparatus to cause a series ofoperational steps to be performed on the computer or other programmableapparatus to produce a computer implemented process such that theinstructions which execute on the computer or other programmableapparatus provide steps for implementing the functions/acts specified inthe block or blocks.

As discussed above, there is a need for effective non-invasive methodsfor determining blood flow distribution. It is believed that none of theexisting methods offer a fully non-invasive cost effective solution tothe problem of flow speed determination. Accordingly, some embodimentsof the inventive concept provide methods, systems and computer programproducts for determining speed distribution of blood flow withoutrequiring the use of dye injection, contrast agent or a contact probe.Some embodiments of the inventive concept use speckle imaging techniquesto determine blood flow distribution. As used herein, a “speckle” imageacquisition refers to the recording of elastically scattered light, i.e.the scattered light has a wavelength that is the same as the incidentlight, from an object illuminated by a coherent light such as the outputfrom a coherent light source. In particular, the “speckle” is actually adiffraction pattern, which is highly correlated to the morphology of theobject being imaged. If certain parts of the object are in translationalmotion, i.e. blood stream flowing in a coronary artery, thecorresponding part or pixels of the speckle image will vary with time infashions different from those parts not undergoing such translationalmotion. This difference in the temporal variation of pixel intensity inthe speckle image provides a mechanism to non-invasively measure flowspeed (m/s or cm/min) in principal vessels. With knowledge of thediameters of the principal vessels that can be determined from the sameset of acquired speckle images, the blood flow rate (ml/min) within FOVmay be determined. The combined information of blood flow rates andtheir distribution in different vessels provide critical data forevaluating the effectiveness of Coronary artery bypass grafting (CABG)and other surgical procedures in improving patients' revascularizationstatus and clinical prognosis. Speckle image acquisition is generallydiscussed, for example, in Velocity measurement of a diffuse object byusing a time-varying speckle by Ohtsubo et al.

Thus, some embodiments of the present inventive concept provide anon-invasive technique for measuring blood flow that provides theability to quantitatively measure blood flow in principal vessels andperfusion distribution in areas perfused by one or more of thoseprincipal vessels as will be discussed further below with respect toFIGS. 1 through 44.

Furthermore, the data acquired from the obtained set of speckle imagescan be verified using flow hemodynamic modeling in accordance with someembodiments of the present inventive concept. In particular, using theNavier-Stokes equation, which provides the governing equation for fluiddynamics, a velocity field associated with the FOV (in the principalvessels) can be obtains. As used herein, “velocity field” refers to adistribution of fluid velocity in space and time. This velocity fieldmay then be used to calculate flow rate as well as other quantities ofinterest, such as pressure. These quantities of interest, for example,flow rates, can then be compared with the experimental data calculatedusing the obtained set of speckle images. Thus, the hemodynamic modelingmay be used to validate the experimental data as well as the success ofthe procedure as will be discuss further below with respect to FIGS. 45through 48.

Referring first to FIG. 1, a non-invasive system for measuring bloodflow in principal vessels of a subject in accordance with someembodiments of the present inventive concept will be discussed. Asdiscussed above, “non-invasive” refers to a system or method that doesnot require the subject to be injected with a dye, penetrated with anobject or touched with an intrabody probe or probes. Thus, as usedherein, the term non-invasive refers to a system or method that makesminimal contact with the subject. As used herein, “subject” refers tothe person or thing being imaged. It will be understood that althoughembodiments of the present inventive concept are discussed herein withrespect to measuring blood flow in principal vessels of the subject,embodiments of the present inventive concept are not limited to thisconfiguration. The subject can be any subject, including a veterinary,cadaver study or human subject. As used herein, “perfusion” refers toblood flow at the tissue perfusion distribution level detected withspeckle imaging.

As illustrated in FIG. 1, the system 100 includes a communicationsdevice 110, a coherent light source unit 120, a camera 130, asynchronization module 170 and an EKG device 180. Although the system ofFIG. 1 is depicted as only including these elements, it will beunderstood that other elements may also be present in the system withoutdeparting from the scope of the present inventive concept. For example,the systems illustrated in the photographs of FIGS. 5 and 21 includeadditional elements not present in the system illustrated in FIG. 1.

Referring again to FIG. 1, in some embodiments, the coherent lightsource unit 120 may be a laser unit, which may include a laser 123 and abeam shaping lens 125. The laser unit 120 may provide a coherent lightsource that illuminates a region of interest 140. The coherent lightsource provided by the laser unit 120 may have a wavelength of fromabout 600 nm to about 1100 nm. As used herein, the “region of interest”refers to the region of the subject that is being imaged, for example,the principal vessels and tissue, organs, etc. to determine blood flowtherein. Although embodiments of the present inventive concept arediscussed primarily herein with respect to blood flow distribution inthe principal vessels, embodiments of the present inventive concept arenot limited to this configuration. For example, blood flow in organs maybe determined without departing from the scope of the present inventiveconcept.

The laser unit 120 may have light output at a fixed or variablewavelength of from about 600 nm to about 1100 nm without departing fromthe scope of the present inventive concept. The laser 120 can beconfigured to illuminate the region of interest 140 with a laser beam127 having substantially constant intensity within FOV of an imagingunit. In some embodiments, the constant or near constant intensity ofthe laser beam can facilitate acquiring speckle images with a highsignal-to-noise (SNR) ratio. The laser 120 can be a low powercontinuous-wave laser. Thus, the subject does not need to wear anyprotective apparatus, for example, clothing or goggles, to shield thesubject from potential adverse effects of the laser. In someembodiments, for example, the laser 120 may be of 633 nm in wavelengthand 1 mW in power.

Use of a laser or other coherent light source having a wavelength offrom about 600 nm to about 1100 nm allows relatively deep penetration oflight into tissue and can provide an accurate determination of bloodflow speed in the principal vessels and the perfusion distribution aswill be discussed further below.

In some embodiments, the laser unit 120 may be used to illuminate thecoronary artery and be triggered by the electrocardiogram (EKG) providedby EKG device 180 through the synchronization module 170 andmeasurements can be taken from the same point outside the heart and thesame point on the heart itself. In other words, the FOV is fixed by twoparameters, the point on the heart and the distance from the cameraoutside the heart. The FOV is kept the same so that the synchronizationcan be performed.

Referring again to FIG. 1, the camera 130 communicates with the laserunit 120 and the communications device 110. The camera 130 is configuredto sequentially acquire at least two speckle images of the region ofinterest during a fixed time period. The faster the camera 130, theshorter the fixed time period has to be for acquiring the same number ofspeckle images. In some embodiments, the camera 130 may be a CCD camera,for example, a Lumenera Lm075 or similar devices.

As used herein, the fixed time period is typically short enough toreduce or possibly minimize motion effects, but long enough to obtainsufficient light signals. Several examples of this fixed time period arediscussed throughout the specification, for example, the fixed timeperiod may be from about 1.0 to about 200 ms, or within a single EKGcardiac cycle. However, it will be understood that the fixed time periodis not limited to the specific time periods discussed herein. Forexample, the fixed time period may be greater than a single EKG cardiaccycle without departing from embodiments discussed herein.

The camera 130 may be configured to acquire from about 50 to about 1000speckle images during the fixed time period. In some embodiments, thecamera may only need to acquire from about 50 to about 500 speckleimages to provide a meaningful result. The fixed time period may beselected based on data associated with in situ determined blood flowspeed. In some embodiments, the fixed time period is relatively short,typically less than 1 second, or from about 1.0 ms to about 200 ms.

The acquisition of the speckle images can be synchronized with themotion of the heart of the subject. For example, in some embodiments,acquisition of the speckle images may be synchronized with the EKG ofthe subject such that the motion of the heart will have minimal effecton determination of blood flow speed. Thus, the fixed time period wouldbe located within a single EKG cardiac cycle.

Referring again to FIG. 1, the communications device 110 is configuredto process the at least two acquired speckle images based on temporalvariation of pixel intensities among the acquired speckle images todetermine spatial distribution of blood flow speed in the principalvessels and perfusion distribution in tissue in the region of interest.The at least two acquired speckle images can be electronically evaluatedand/or processed using an image processing algorithm that combinestemporal and spatial calculations of the at least two acquired speckleimages. The at least two acquired speckle images have a directrelationship to the blood flow speed in the principal vessels and theperfusion distribution.

In particular, some embodiments of the present inventive concept usespeckle imaging techniques to yield blood flow speed in the principalvessels and perfusion distribution over the FOV. As used herein, FOVrefers to the area of the imaged object that can be viewed by theimaging sensor. Due to the coherence among the scattered light fromdifferent parts of the illuminated region of the imaged object, theintensity of the scattered light arriving at a detecting element of animaging sensor depends on the relative spatial relation among thedifferent parts. The dependency leads to a “speckle” appearance of theacquired image since intensity of scattered light having an opticalwavelength of from about 200 nm to about 2000 nm can vary quickly over asmall spatial domain with a size of about 10 cm. These concepts will bediscussed further below.

Referring now to FIGS. 2A and 2B, a data processing system 200 that maybe used in the system 100 illustrated in FIG. 1 in accordance with someembodiments of the inventive concept will be discussed. The dataprocessing system 200 may be included in the communications device 110,the camera 130 or split between various elements of the system 100without departing from the scope of the present inventive concept. Asillustrated in FIG. 2, an exemplary embodiment of a data processingsystem 200 suitable for use in the system 100 of FIG. 1 includes a userinterface 244 such as a keyboard, keypad, touchpad or the like, I/O dataports 246 and a memory 236 that communicates with a processor 238. TheI/O data ports 246 can be used to transfer information between the dataprocessing system 200 and another computer system or a network. Thesecomponents may be conventional components, such as those used in manyconventional data processing systems, which may be configured to operateas described herein.

Referring now to FIG. 2B, a more detailed block diagram of the dataprocessing system 200 in accordance with some embodiments of the presentinventive concept will be discussed. The processor 238 communicates witha display 345 via and address/data bus 347, the memory 236 via anaddress/data bus 348 and the I/O data ports 246 via an address/date bus349. The processor 238 can be any commercially available or custommicroprocessor or ASICs. The memory 236 is representative of the overallhierarchy of memory devices containing the software and data used toimplement the functionality of the data processing system 200. Thememory 236 can include, but is not limited to, the following types ofdevices: cache, ROM, PROM, EPROM, EEPROM, flash memory, SRAM, and DRAM.

As shown in FIG. 2B, the memory 236 may include several categories ofsoftware and data used in the data processing system 200: an operatingsystem 352; application programs 354; input/output (I/O) device drivers358; and data 356. As will be appreciated by those of skill in the art,the operating system 352 may be any operating system suitable for usewith a data processing system, such as OS/2, AIX or zOS fromInternational Business Machines Corporation, Armonk, N.Y., Windows95,Windows98, Windows2000, WindowsXP, or Vista from Microsoft Corporation,Redmond, Wash., Unix, Linux, LabView, or a real-time operating systemsuch as QNX or VxWorks, or the like. The I/O device drivers 358typically include software routines accessed through the operatingsystem 352 by the application programs 354 to communicate with devicessuch as the I/O data port(s) 246 and certain memory 236 components. Theapplication programs 354 are illustrative of the programs that implementthe various features of the data processing system 200 included a systemin accordance with some embodiments of the present inventive concept andpreferably include at least one application that supports operationsaccording to some embodiments of the present inventive concept. Finally,the data 356 represents the static and dynamic data used by theapplication programs 354, the operating system 352, the I/O devicedrivers 358, and other software programs that may reside in the memory236.

As illustrated in FIG. 2B, the data 356 according to some embodiments ofthe present inventive concept may include acquired speckle images 360,intermediate data 361, calculated blood flow rates 363 and modeling data364. Although the data 356 illustrated in FIG. 2B includes threedifferent files 360, 361, 363 and 364, embodiments of the presentinventive concept are not limited to this configuration. Two or morefiles may be combined to make a single file; a single file may be splitinto two or more files and the like without departing from the scope ofthe present inventive concept.

As further illustrated in FIG. 2B, the application programs 354 mayinclude a light source trigger module 351, an image capture module 352,a processing module 353 and a modeling module 354 in accordance withsome embodiments of the inventive concept. While the present inventiveconcept is illustrated, for example, with reference to the light sourcetrigger module 351, the image capture module 352, the processing module353 and the modeling module 354 being application programs in FIG. 2B,as will be appreciated by those of skill in the art, otherconfigurations may also be utilized while still benefiting from theteachings of the present inventive concept. For example, the lightsource trigger module 351, the image capture module 352 the processingmodule 353 and the modeling module 354 may also be incorporated into theoperating system 352 or other such logical division of the dataprocessing system 300. Thus, the present inventive concept should not beconstrued as limited to the configuration of FIG. 2B, but is intended toencompass any configuration capable of carrying out the operationsdescribed herein.

Furthermore, while the light source trigger module 351, the imagecapture module 352 the processing module 353 and the modeling module 354are illustrated in a single data processing system, as will beappreciated by those of skill in the art, such functionality may bedistributed across one or more data processing systems. Thus, thepresent inventive concept should not be construed as limited to theconfiguration illustrated in FIGS. 2A and 2B, but may be provided byother arrangements and/or divisions of function between data processingsystems.

In particular, the light source trigger module 351 may be configured toilluminate a region of interest with a coherent light source. Thecoherent light source may have a wavelength of from about 600 nm toabout 1100 nm as discussed above. The image capture module 352 may beconfigured to sequentially acquire at least two speckle images of theregion of interest during a fixed time period. The processing module 353may be configured to process the at least two acquired speckle imagesbased on a diffraction pattern of each the at least two speckle imagesto determine spatial distribution of blood flow speed in the principalvessels and perfusion distribution in tissue in the region of interest.

The modeling module 354 may be configured to calculate a velocity fieldfor the region of interest; calculate blood flow speed in the region ofinterest based on the calculated velocity field; and compare thecalculated blood flow in the region of interest to the blood flow speeddetermined using the acquired at least two speckle images of the regionof interest to verify results obtained using the at least two speckleimages. In some embodiments, the modeling module 354 is configured tocalculate the velocity field using Equations 9 and 10 set out below.

Thus, blood flow speed as well as other quantities may be calculatedusing both the speckle method and the velocity field method before aprocedure is performed on a subject and after a procedure is performedon the subject to verify that the procedure was successful. By comparingthe measurements/quantities before and after the procedure, the successof the procedure may be determined, which will be discussed furtherbelow.

Referring now to the flowcharts of FIGS. 3 and 4, operations of anon-invasive method for measuring blood flow in principal vessels of asubject will be discussed. As illustrated in FIG. 3, operations begin atblock 315 by illuminating a region of interest in the heart with acoherent light source. The coherent light source may have a wavelengthof from about 600 nm to about 1100 nm. Providing a coherent light sourcewith a wavelength of from about 600 nm to about 1100 nm may allow fornon-invasive, deep penetration of light into tissues and provides anaccurate determination of blood flow speed in the principal vessels andthe perfusion distribution within the layer of light penetration.

In some embodiments, the coherent light source may be provided by alaser configured to illuminate the region of interest. The laser mayhave a fixed or variable wavelength. The laser may produce a beam havingsubstantially constant intensity within a FOV of an imaging unit. Thelaser may be a low energy and continuous-wave laser such that thesubject does not require any protective apparatus to shield the subjectfrom effects of the laser.

Referring again to FIG. 3, operations continue at block 325 bysequentially acquiring at least two speckle images of the region ofinterest during a fixed time period. The fixed time period may beselected based on data associated with in situ determined blood flowspeed. In some embodiments, the at least two speckle images may beacquired in synchronization with motion of a heart of the subject suchthat the motion of the heart will have minimal effect on determinationof blood flow speed using the acquired at least two speckle images. Forexample, the fixed time period can correspond to a single EKG cardiaccycle or defined portion thereof cycle.

The camera may be configured to acquire the at least two speckle imagesduring the fixed time period. In some embodiments, from about 50 toabout 1000 speckle images may be acquired using the camera during thefixed time period of from about 1 ms to about 200 ms. In someembodiments, about 200 to about 500 speckle images may be acquired.Higher numbers of speckle images typically allow better signal-to-noiseratios in the calculated LSCI image but take longer time to acquire.

Referring again to FIG. 3, operations continue at block 335 byelectronically processing the acquired speckle images based on thetemporal variation of the pixel intensities in the acquired speckleimages to generate a laser speckle contrast imaging (LSCI) image anddetermine spatial distribution of blood flow speed in the principalvessels and perfusion distribution in tissue in the region of interestfrom the LSCI image.

In some embodiments, electronically evaluating speckle image data mayinclude electronically evaluating the acquired speckle images using animage processing algorithm that combines temporal and spatialcalculations of the acquired speckle images to generate a LSCI image anddetermine spatial distribution of blood flow speed. The at least twospeckle images may have a direct relationship to the blood flow speed inthe principal vessels and the perfusion distribution which are utilizedin generating an LSCI image for determination of the spatialdistribution of blood flow speed. For example, following equations canbe used to obtain the intensity at each pixel of the LSCI image K (i, j)from the acquired speckle image set {I_(n)} with n=1, 2, . . . , N,i.e.,

$\begin{matrix}{{{K\left( {i,j} \right)} = \frac{\sigma \left( {i,j} \right)}{\mu \left( {i,j} \right)}},{where}} & {{Equation}\mspace{14mu} (1)} \\{{{\mu \left( {i,j} \right)} = {\frac{1}{N}{\sum\limits_{n = 1}^{N}{I_{n}\left( {i,j} \right)}}}},} & {{Equation}\mspace{14mu} (2)} \\{{\sigma \left( {i,j} \right)} = {\sqrt{\frac{1}{N}{\sum\limits_{n = 1}^{N}\left( {{I_{n}\left( {i,j} \right)} - {\mu \left( {i,j} \right)}} \right)^{2}}}.}} & {{Equation}\mspace{14mu} (3)}\end{matrix}$

In the above calculations, I_(n)(i, j) refers to the pixel at (i, j)location in a speckle image acquired at nth time point and N (>1) is thetotal number of acquired speckle images.

It will be understood that the operations of blocks 315, 325 and 335 maybe performed before and after a procedure performed on the subject. Theresults before and after the procedure may be compared to verify thesuccess of the procedure in the subject.

Referring now to FIG. 4, operations for a non-invasive method formeasuring blood flow in principal vessels of a subject in accordancewith some embodiments will be discussed. Operations begin at block 415by illuminating a region of interest in the heart with a coherent lightsource, wherein the coherent light source has a wavelength of from about600 nm to about 1100 nm. At least two speckle images of the region ofinterest are sequentially acquired during a fixed time period (block425) in synchronization with the motion of the heart. Temporal andspatial variation of pixel intensities of the at least two acquiredspeckle images are electrically evaluated to determine spatialdistribution of blood flow speed in the principal vessels and perfusiondistribution in tissue in the region of interest of the heart (block435).

A velocity field for the region of interest in the heart is calculated(block 445). In some embodiments, the velocity field is calculated usingequations (9) and (10) set out below. Blood flow speed in the region ofinterest of the heart based on the calculated velocity field iscalculated (block 455). The calculated blood flow speed in the region ofinterest in the heart is compared to the blood flow speed determinedusing the acquired at least two speckle images of the region of interestto verify results obtained using the at least two speckle images (block465). Thus, embodiments of the present inventive concept may be used toverify experimental results as will be discussed further below.

It will be understood that the operations of blocks 415, 425, 435, 445,455 and 465 may be performed before and after a procedure performed onthe subject. The results before and after the procedure may be comparedto verify the success of the procedure in the subject.

The following non-limiting examples are provided by way of example.

EXAMPLES

Referring to FIG. 5, a digital photograph of a prototype system 500 todetect flow speed using laser speckle contrast imaging (LSCI) technologyin accordance with some embodiments of the present inventive conceptwill be discussed. As illustrated in FIG. 5, the system includes acommunication device 510, such as a laptop computer, a laser unit 520including a laser generator and a focusing lens, a camera 530, a flowgenerator 580, flow liquid 590 and a flow target 585. Table 1 set outbelow summarizes the actual equipment/devices used in this experiment.

TABLE 1 Devices used in Experiment 1 Notes CCD camera (530) LumeneraLm075 Laser (520) 633 nm in wavelength, 1 mW in power Liquid used inflow (590) 20% Intralipid 1 to 4 ratio mixed with water plus fruit colorComputer/Communications Laptop PC Device (510)

As discussed above, the faster the camera 530, the smaller the fixedtime period has to be to obtain an adequate number of speckle images toprovide a meaningful result. Thus, the limitation of the frame rate ofthe camera 530 in the prototype system may have impacted the finalresult of this experiment. The laser 520 is a low power continuous-wavelaser providing a single-wavelength coherent light source. Thus, thesubject of the imaging does not typically require any protection fromsuch a laser, such as protective clothing or eyewear. The laser 520produces a beam having a wavelength of from about 600 nm to about 1100nm in some embodiments. During the experiment, the laser beam producedby the laser 520 is used to illuminate the region of interest withsubstantially constant intensity with the FOV of the imaging unit. Thisis an important aspect of the experiment because it allows the resultingimages to have a high SNR.

Colored intralipid was used as the flow liquid 590 during the experimentdue to the fact that a light scattering characteristics of the coloredintralipid is similar to those of mammalian blood. Thus, the coloredintralipid mimics the blood flowing in the human body. Thecommunications device 510 used was a laptop computer, althoughembodiments of the present inventive concept are not limited to the useof a laptop computer. The acquired speckle images are provided to thecommunications device 510 and are used to calculate blood flow inaccording with some embodiments of the present inventive concept. Asdiscussed above, the data is calculated using an image processingalgorithm that combines temporal and spatial calculations of theacquired speckle images. Thus, spatial distribution of blood flow speedin principal vessels and perfusion distribution can be determined.

Referring now to FIGS. 6 through 9, close up photographs of the devicesillustrated in FIG. 5 used during the first experiment are provided. Inparticular, FIG. 6 is a close up photograph of the laser unit 520; FIG.7 is a close up photograph of the camera 530; FIG. 8 is a close upphotograph of the communications device 510; and FIG. 9 is a close ofthe flow liquid 590.

Table 2 set out below summarizes parameters for the camera 530 usedduring the first experiment. The parameters are for the camera 530 whilethe image sequence is acquired.

TABLE 2 Length of image sequence ~1 second Frame rate ~95 frames/secondImage resolution 320 * 240 pixels Exposure time per frame 3 ms Gain 1

The upper limit (V_(limit)) of flow that can be detected based on thesetup of the first experiment can be summarized by the followingequation:

$\begin{matrix}{V_{limit} = \frac{\Delta \; L}{10\tau}} & {{Equation}\mspace{14mu} (4)}\end{matrix}$

In Equation 1, ΔL is the diameter of the tube, which was 0.26 cm in thefirst experiment;

$\tau = {{0.4T} = \frac{0.4}{f}}$

is the estimated exposure time, 3.0 ms in the first experiment; and f isthe frame rate. Thus, in the first experiment V_(limit) can be roughlyestimated at about 9.0 cm/second.

Referring now to FIGS. 10 and 11, photographs of the flow generationsystem used in the first experiment will be discussed. FIG. 10illustrates the flow generation system 580. The higher the bottle in theflow generation system 580, the faster the flow of the flow liquid 590.FIG. 11 is a close up photograph of the tube target 585 used as the flowtarget for the first experiment. Table 3 below summarizes therelationship between the change in height of the bottle and the flow ofliquid in the flow generation system in accordance with the firstexperiment. The change in height is measured from the height of thebottle to the end of the tube.

TABLE 3 Time Volume delta height flow rate flow rate predicted flow(secs) (cm³) (cm) (cm³/sec) (ml/min) rate (ml/min) 68 100 108 1.5 88.290.6 73 100 98 1.4 82.2 83.0 80 100 88 1.3 75.0 75.4 90 100 78 1.1 66.767.6 103 100 68 1.0 58.3 59.8 120 100 58 0.8 50.0 51.8 136 100 48 0.744.1 43.7 173 100 38 0.6 34.7 35.4

Thus, as illustrated by the delta height (cm) and flow columns of Table3, the higher the bottle, the faster the flow rate of the coloredintralipid liquid.

The estimated relationship between delta height (cm) and flow rate(ml/min) is represented by Equation (2) set out below:

$\begin{matrix}{{{{flow}\mspace{14mu} {rate}} = {1.34 \times \sqrt[0.9]{\Delta \; h}}};} & {{Equation}\mspace{14mu} (5)}\end{matrix}$

Where Δh is the change in height of the bottle relative to the verticalposition of the tube end. FIG. 12 summarizes the data in Table 3 and isa graph illustrating delta height (cm) vs. flow rate (ml/min).

Four cases of flow rate were measured by the LSCI method in the firstexperiment setup, no flow, flow 1, flow 2 and flow 3. The details ofeach of the flow rates are summarized in Table 4 set out below. Thefirst experiment was repeated three times for each case to ensureaccuracy and repeatability. FIGS. 13A through 13D illustrate resultantimages obtained for the “no flow” state. FIG. 13A is an averaged imageobtained by averaging in a pixel-to-pixel fashion 97 frames for the “noflow” case; FIG. 13B is a vertical line profile image in the middle ofinverted speckle contrast image for the “no flow” case; FIG. 13C is aninverted speckle contrast image for the “no flow” case; and FIG. 13D isa colorized inverted speckle contrast image for the “no flow” case.

FIGS. 14A through 14D illustrate resultant images obtained for the “flow1” case. FIG. 14A is an averaged image obtained by averaging in apixel-to-pixel fashion 97 frames for the “flow 1” case; FIG. 14B is avertical line profile image in the middle of the inverted specklecontrast image for the “flow 1” case; FIG. 14C is an inverted specklecontrast image for the “flow 1” case; and FIG. 14D is a colorizedinverted speckle contrast image for the “flow 1” case.

FIGS. 15A through 15D illustrate resultant images obtained for the “flow2” case. FIG. 15A is an averaged image obtained by averaging in apixel-to-pixel fashion 89 frames for the “flow 2” case; FIG. 15B is avertical line profile image in the middle of an inverted specklecontrast image for the “flow 2” case; FIG. 15C is an inverted specklecontrast image for the “flow 2” case; and FIG. 15D is a colorizedinverted speckle contrast image for the “flow 2” case.

FIGS. 16A through 16D illustrate resultant images obtained for the “flow3” case. FIG. 16A is an averaged image obtained by averaging in apixel-to-pixel fashion 89 frames for the “flow 3” case; FIG. 16B is avertical line profile image in the middle of an inverted specklecontrast image for the “flow 3” case; FIG. 16C is an inverted specklecontrast image for the “flow 3” case; and FIG. 16D is a colorizedinverted speckle contrast image for the “flow 3” case.

FIGS. 17A through 17D are the averaged images for each of the cases, “noflow”, “flow 1”, “flow 2”, and “flow 3,” respectively. FIGS. 18A through18D are colorized inverted speckle contrast images for each of thecases, “no flow”, “flow 1”, “flow 2”, and “flow 3,” respectively. FIGS.19A through 19D are vertical line profile images for each of the cases,“no flow”, “flow 1”, “flow 2”, and “flow 3,” respectively.

Table 4 set out below, summarizes the data for all four flow cases. Inparticular, the relationship between delta height and predicted flow isreadily apparent. FIG. 20 is a graph illustrating predicted flow rate(ml/min) vs. inverted speckle contrast image pixel intensity as set outin Table 4.

TABLE 4 delta inverted speckle height predicted flow contrast imageStatus (cm) rate (ml/min) pixel intensity No flow 0 0 15 Flow 1 10 10.667 Flow 2 100 84.5 88 Flow 3 193 150.7 108

To summarize the first experiment, the laser speckle contrast imagingsetup is clearly able to differentiate between a no flow state and thethree flow speed cases. However, the sensitivity and precision were notideal and the point of “no flow” was not consistent with the other threeflow points as illustrated in FIG. 20. Some of the imprecision may bedue to using a bottle for the flow speed generation method. This mayhave caused variation and lack of constant flow. Furthermore, frame rateof the camera may have limited to number of speckle images that could beobtained. The laser beam intensity used during the experiment was unevenin the FOV, i.e. there are dark spots in the FOV.

However, even given this imprecision, the results shown that the higherthe frame rate the better the LSCI image quality and it was discoveredthat the exposure time should be as long as it can in the condition thatthe same frame rate can be achieved.

Referring first to FIG. 21, a photograph of the system 2100 for a secondexperiment to detect flow speed using the LSCI technology in accordancewith some embodiments of the present inventive concept will bediscussed. As illustrated in FIG. 21, the system includes acommunications device 2110, such as a laptop computer, a laser unit 2120including a laser generator and a beam shaping lens, a camera 2130, aflow generator 2181 provided by a biomedical pump, flow liquid 2190, aflow target 2185 and an electromagnetic flow detector 2191. Table 5 setout below summarizes the actual equipment/devices used in thisexperiment.

TABLE 5 Devices used in Experiment 2 Notes CCD camera (2130) LumeneraLm075 Laser (2120) 633 nm in wavelength, 1 mW in power Liquid used inflow (2190) 20% Intralipid Saline water 0.9% Biomedical pump (2181) Withelectromagnetic flow detector (2191) Communications Device/ Laptop PCComputer(2110)

As discussed above, the faster the camera 2130, the smaller the fixedtime period has to be to obtain an adequate number of speckle images toprovide a meaningful result. Thus, the limitation of the frame rate onthe camera 2130 may have impacted the final result of this experiment.The laser 2120 is a low power laser providing a single coherent lightsource. Thus, the subject of the imaging does not typically require anyprotection from the laser, such as protective clothing or eyewear. Thelaser 2120 produces a beam having a wavelength of from about 600 nm toabout 1100 nm in some embodiments. During the experiment, the laser beamproduced by the laser 2120 is used to illuminate the region of interestwith substantially constant intensity with the FOV of the imaging unit.This may allow the speckle contrast images to have a highsignal-to-noise ratio.

Colored intralipid was used as the flow liquid 2190 during theexperiment due to the fact that light scattering characteristics ofintralipid are similar to those of mammalian blood. Thus, the coloredintralipid will mimic the blood flowing in the human body. Thecommunications device 2110 used was a laptop computer, althoughembodiments of the present inventive concept are not limited to the useof a laptop computer. The acquired speckle images are provided to thecommunications device 2110 and are used to calculate blood flow inaccording with some embodiments of the present inventive concept. Asdiscussed above, the data are calculated using an image processingalgorithm that combines temporal and spatial calculations of theacquired speckle images. Thus, spatial distribution of blood flow speedin principal vessels and perfusion distribution can be determined.

Referring now to FIGS. 22 through 27, close up photographs of thedevices illustrated in FIG. 21 used during the second experiment areprovided. In particular, FIG. 22 is a close up photograph of the laserunit 2120; FIG. 23 is a close up photograph of the camera 2130; FIG. 24is a close up photograph of the communications device 2110; FIG. 25 is aclose up photograph of the biomedical pump 2181; FIG. 26 is a close upphotograph of the electromagnetic flow detector 2191; and FIG. 27 is aclose up photograph of the flow target 2185.

Table 6 set out below summarizes parameters for the camera 530 usedduring the second experiment. The parameters are for the camera 2130while the image sequence is acquired.

TABLE 6 Length of image sequence ~1 second Frame rate ~95 frames/secondImage resolution 320 * 240 Exposure time per frame 3 ms (split into 3part to calculate std and average every 3 continuous frames) Gain 1Working distance ~1.5 m Aperture 2~4 Length of the video loop 3 * 1second

Estimation of the blood flow rate and speed in accordance with thesecond experiment will be discussed. Since the LSCI technology measuresthe flow speed (cm/min) rather than flow rate (ml/min), the range of theflow speed of blood in the main branches of coronary artery can beestablished. The second experiment focused on this range. It iscontemplated that the flow rate can be calculated using the flow speedand a cross sectional area of the vessel at the same location. Table 7set out below illustrates the estimated blood flow rate and speed rangein LAD.

TABLE 7 Average diameter flow rate range flow speed range of LAD (cm)(ml/min, cm³/min) (cm/min) 0.4 0~100 0~796

Procedures used for in accordance with the second experiment will now bediscussed. The biomedical pump was calibrated by comparing measuredliquid volume during a certain time with the reading from theelectromagnetic flow detector 2591 as illustrated in Table 8 set outbelow. In particular, pump calibration using 20% Intralipid solutionmixed by saline water with 1:4 ratio (volume was measured from 100 mL to300 mL and reading from the detector was recorded when liquid reaches200 mL).

TABLE 8 flow rate flow rate reading reading Calculated flow Time TimeVolume from pump from pump flow rate Percentage speed (s) (min) (mL)(L/min) (mL/min) (ml/min) Error (cm/min) 15 0.25 200 0.78 780 800 3%1095 18 0.30 200 0.65 650 667 3% 913 22 0.37 200 0.55 550 545 1% 772 260.43 200 0.45 450 462 3% 632 30 0.50 200 0.39 390 400 3% 548 49 0.82 2000.24 240 245 2% 337 62 1.03 200 0.18 180 194 7% 253 111 1.85 200 0.11110 108 2% 154

Three Intralipid solutions with different concentrations were measuredto simulate light scattering characteristics of blood as will bediscussed below and to examine the stability of the LSCI technology. Thethree intralipid concentrations are summarized below in Table 9. Table10 summarizes the volume and linear flow rate range measured during thesecond experiment.

TABLE 9 Ratio of 20% Intralipid to Intralipid saline water concentrationrepeat Note 1:39 0.5%   3 Tube surface is processed by sand paper andcollimated reflectance lights were greatly reduced. Aperture of thecamera was large. ~2.5 1:19 1% 3 Tube surface is processed by sand paperand collimated reflectance lights were greatly reduced. Aperture of thecamera was medium. ~3 1:9 2% 3 Same as above

TABLE 10 Estimated flow rate reading Corresponding flow maximum in ⅜inch Calculated rate in 4 mm flow rate in diameter tube flow speeddiameter LAD coronary artery 0~1000 mL/min 0~1404 cm/min 0~176 mL/min100 mL/min

Results of the second experiment will now be discussed. Table 11 set outbelow summarizes the results for (1): 0.5% Intralipid solution (1:39);(2): 1% Intralipid solution (1:19); (3) 2% Intralipid solution (1:9) inaccordance with the second experiment.

TABLE 11 (Std is standard deviation, L is the inversed speckle contrastsee Equation 6 for details) flow rate reading flow Corresponding frompump speed LAD flow rate Std of L Std of L Std of L (mL/min) (cm/min)(mL/min) L (1) (1) L (2) (2) L (3) (3) 0 0 0 56 0.6 51 0.5 44 0.1 100140 18 88 4.7 88 1.0 75 2.0 200 281 35 97 4.6 94 2.0 92 1.0 300 421 53105 4.3 110 2.1 104 1.1 400 562 71 111 5.5 112 0.9 109 0.3 500 702 88113 2.9 117 2.8 117 1.4 600 842 106 121 4.0 118 2.2 121 2.8 700 983 123120 1.0 113 3.4 127 0.3 800 1123 141 115 5.6 109 3.1 128 0.7 900 1264159 113 3.8 113 2.1 123 5.7 1000 1404 176 112 3.8 106 6.2 124 3.8

Referring now to FIG. 28, flow speed and corresponding LAD flow rate vs.the inverted speckle contrast image pixel intensity 1/K; Resultsobtained with ratio of 20% Intralipid to saline water of (1) 1:39; (2)1:19; (3) 1:9 will be discussed. As illustrated in FIG. 28, the morediluted Intralipid solution has a larger background noise while flowreading is zero; after the flow speed exceeds 800 cm/min, the 1/K beginsto saturate and has become less sensitive to the flow speed change; andcompared to the huge concentration rage of the Intralipid solution,there is only very small change in the curves.

The quantitative relation between laser speckle contrast image pixelintensity and the flow speed will now be discussed. To make a positivecorrelation between the flow speed and the calculated image pixelintensity, the following equation is used to construct the invertedspeckle contrast image L:

$\begin{matrix}{{L\left( {i,j} \right)} = {\frac{1}{K\left( {i,j} \right)} = \frac{\mu \left( {i,j} \right)}{\sigma \left( {i,j} \right)}}} & {{Equation}\mspace{14mu} (6)}\end{matrix}$

The quantitative relationship between laser speckle contrast image pixelK(i,j) and the flow speed have been derived as the following

K(i,j)∝√{square root over (τ_(c)(i,j)/T)}  Equation (7)

where T is the camera integration time, τ_(c) is a correlation time ofscattering particles undergoing motion of speed v and given by

${\tau_{c} = \frac{\lambda}{2\pi \; v}},$

λ is laser light wavelength. Based on the above relation, the invertedspeckle image pixel intensity can be written as

L(i,j)=L ₀ +a√{square root over (v(i,j))}  Equation (8)

where L₀ is an added term to account for background noise and should bezero after the baseline has been removed; a is a constant related to theimaging parameters, laser parameters, time/spatial smoothing parametersfor obtaining K and the components of the liquid. Thus, after removingthe baseline from the data of “zero flow” case from each singlemeasurements in case (1)˜(3), the Equation (8) can be used to fit themeasured data.

FIG. 29 is a graph illustrating flow speed vs. the L with curve fittingbased on Equation (8). Table 12 summarizes the results of curve fitting,with R as the value of confidence of fitting (R=1 means perfectfitting).

TABLE 12 Case (1) case (2) case (3) L₀ 0 0 0 a 2.3149 2.5413 2.7606 R0.9906 0.9797 0.9974

Based on the prediction model (curve fitting), flow speed inmeasurements (1)˜(3) were obtained to compare with the flow speedcalculated from the pump readings. The results are depicted in FIG. 30,a graph illustrating the average of calculated flow speed (cm/min) fromL in case (1)˜(3) vs. the flow speed determined from the pump readings.

To summarize the second experiment, the experiment showed that theinverted speckle image pixel intensity L acquired with the currentimaging system can correlate to the speed of the flow within certainrange, (flow speed <800 cm/min or LAD flow rate<100 mL/min), whichcorresponds to the maximum blood flow rate in average main branch ofcoronary artery. Compared to the huge concentration range of theIntralipid solution, there is only very small change in the curves. Thelight scattering parameters of blood might fall in between case (1) and(3) as illustrated in Table 13 set out below, where μ_(a) is absorptioncoefficient, μ_(s) is scattering coefficient, g is anisotropy factor andμ_(s)′=(1−g)μ_(s) is the reduced scattering coefficient.

TABLE 13 oxygenated λ = 633 nm blood 10% Intralipid u_(a) (1/mm) 0.20.0003 u_(s) (1/mm) 31 34.75 g 0.99 0.83 u_(s)′ (1/mm) 0.31 5.98

When bubbles are trapped in the biomedical pump, they may affect theaccuracy of the flow reading from electromagnetic flow meter and mightaffect results of measurement. Specular reflectance from the smoothsurface of the tube will cause saturation of the camera and overwhelmthe speckle information as shown in FIGS. 31A through 31D. In the secondexperiment, sand paper was used to create rough surface on the tube toeliminate this influence, the results of which are illustrated in FIGS.32A through 32D, i.e. specular reflectance is removed by roughening thesurface of the tube. The speed detection limit of L depends on the framerate and the number of frames used to generate the speckle contrastimage somehow is related to the signal-to-noise ratio of L. Smoothing ofthe original images with a moving window of 3 to 5 frames helps increasethe signal-to-noise ratio.

Further experiments performed in accordance with some embodiments of thepresent inventive concept will now be discussed with respect to FIGS. 33through 43. FIG. 33 is a digital photograph of a prototype system setupused to detect blood flow speed using the LSCI technology in accordancewith some embodiments of the present inventive concept. The experimentdiscussed with respect to FIGS. 33 through 43 is similar to theexperiment discussed above with respect to FIGS. 21 through 32, but anextra reservoir 3397 is present to add red blood cells into thecirculation from a blood bag 3398 (expired and for research use only),which is acquired from the Red Cross. Thus, many of the details of theexperiment will not be repeated herein.

Table 14 set out below summarizes the actual equipment/devices used inthis experiment.

TABLE 14 Device Parameters CCD camera Lumenera Lm075 Laser 633 nminwavelength, 1 mW in power Liquid used in Blood (1 unit of red bloodcells with flow hematocrit 70% + 1000 mL saline water) Saline water 0.9%Biomedical pump With electromagnetic flow rate detector Computer LaptopPC

Table 15 set out below summarizes parameters for the camera used duringthe second experiment. The parameters are for the camera while the imagesequence is acquired.

TABLE 15 Length of image sequence 1 second Frame rate ~95 frames/secondImage resolution 320 * 240 Exposure time per frame 3 ms Gain 1 workingdistance 1.5~4 m Aperture 2 Length of the video loop 3 * 1 second (splitinto 3 part to calculate std and average every 3 continuous frames)

Four flow cases were measured by LSCI using the setup in FIG. 33 andeach situation was performed three times to test the repeatability. Theflow cases tested are “tube is clamped”, “pump is reading 100 mL”, “pumpis reading 500 mL” and “pump is reading 1000 mL” Table 16 set out belowsummarized the blood flows measured and the results for each situation.

TABLE 16 flow rate reading Corresponding from pump flow speed LAD flowrate L std of L (mL/min) (cm/min) (mL/min) (blood) (blood) Tube clamped35 2 0 0 0 84 1 100 140 18 144 2 200 281 35 153 4 300 421 53 156 6 400562 71 164 6 500 702 88 168 4 600 842 106 174 3 700 983 123 179 3 8001123 141 180 10 900 1264 159 188 1 1000 1404 176 191 4 (Std is standarddeviation, L is the inversed speckle contrast see Equation 6 fordetails)

FIGS. 34A through 34D illustrate resultant images obtained for the “tubeis clamped” case. FIG. 34A is an averaged image of 90 frames for the“tube is clamped” case; FIG. 34B is a vertical line profile image in themiddle of inverted speckle contrast image for the “tube is clamped”case; FIG. 34C is an inverted speckle contrast image for the “tube isclamped” case; and FIG. 34D is a false-color inverted speckle contrastimage for the “tube is clamped” case.

FIGS. 35A through 35D illustrate resultant images obtained for the “pumpreads 100 mL” case. FIG. 35A is an averaged image of 97 frames for the“100 mL” case; FIG. 35B is a vertical line profile in the middle of aninverted speckle contrast image for the “100 mL” case; FIG. 35C is a ninverted speckle contrast image for the “100 mL” case; and FIG. 35D is afalse-color inverted speckle contrast image for the “100 mL” case.

FIGS. 36A through 36D illustrate resultant images obtained for the “pumpreads 500 mL” case. FIG. 36A is an averaged image of 97 frames for the“500 mL” case; FIG. 36B is a vertical line profile image in the middleof an inverted speckle contrast image for the “500 mL” case; FIG. 36C isa n inverted speckle contrast image for the “500 mL” case; and FIG. 36Dis a false-color speckle contrast image for the “500 mL” case.

FIGS. 37A through 37D illustrate resultant images obtained for the “pumpreads 1000 mL” case. FIG. 37A is an averaged image of 98 frames for the“1000 mL” case; FIG. 37B is a vertical line profile image in middle ofan inverted speckle contrast image for the “1000 mL” case; FIG. 37C isan inverted speckle contrast image for the “1000 mL” case; and FIG. 37Dis a false-color inverted speckle contrast image for the “1000 mL” case.

FIGS. 38A through 38D are averaged images for each of the cases, “pumpclamped”, “100 mL”, “500 mL”, and “1000 mL,” respectively. FIGS. 39Athrough 39D are colorized inverted speckle contrast images for each ofthe cases, “pump clamped”, “100 mL”, “500 mL”, and “1000 mL,”respectively. FIGS. 40A through 40D are vertical line profile images foreach of the cases, “pump clamped”, “100 mL”, “500 mL”, and “1000 mL,”respectively.

FIG. 41 of a graph of flow speed vs. L with to the line fitting the databased on Equation (8). Table 17 set out below summarized the curvefitting parameters using Equation (8).

TABLE 17 L (1) L₀ 100 a 3.0 R 0.96

The laser speckle imaging setup illustrated in FIG. 33 is clearly ableto differentiate different flow speeds of human blood. In particular,when the reading from the pump is zero, the LSCI image still hascontrast related to flow speed as illustrated in FIGS. 43A through 43D.Until the tube is clamped, the contrast disappears as shown in FIGS. 44Athrough 44D. Thus, when the flow speed is very low, is the pump readingstill accurate? The power of the speed term v in Equation (8) aftercurve fitting was found not equal to 0.5, which is different from theIntralipid solution as discussed above with respect to FIGS. 21-32. Ifthe data related to “no flow” case is discarded, one can see that therest of data fit the Equation (8) well, the result is summarized in thegraph of FIG. 42 illustrating flow speed vs. L with Equation (8) afterthe baseline removed. Thus, the Equation (6) with the power factor of0.5 will fit the data well. Table 18 set out below summarizes the curvefitting parameters.

TABLE 18 L (1) L₀ 120 a 1.86 R 0.994

As discussed above, the data obtained using the speckle images discussedabove can be verified using hemodynamic modeling as will be discussedwith respect to FIGS. 45 through 47. The Navier-Stokes equation providesthe governing equation for fluid dynamics.

The Navier-Stokes equation is set out below in Equations (9) and (10):

$\begin{matrix}{{\rho \cdot \left( {\frac{\partial u}{\partial t} + {u{\nabla{\cdot u}}}} \right)} = {{- {\nabla p}} + {\mu \cdot {\nabla^{2}u}} + F}} & {{Equation}\mspace{14mu} (9)} \\{{\frac{\partial\rho}{\partial t} + {\nabla{\cdot \left( {\rho \; u} \right)}}} = 0} & {{Equation}\mspace{14mu} (10)}\end{matrix}$

where ρ is the density (kg/m³), u is the velocity vector (m/s), p is thepressure (m/s), F is the volume force vector (N/m³) and m is theviscosity.

Solving the Navier-Stokes equations produces a velocity field, i.e. adistribution of fluid velocity in space and time. Once this velocityfield is obtained, other quantities of interest, such as flow rate anddrag force, can be calculated. These calculated quantities can then becompared to the experimental data obtained using the speckle imagesdiscussed above to validate the data.

Furthermore, these quantities may be calculated before a procedure isperformed on a subject and after a procedure is performed on the subjectto verify that the procedure was successful. For example, measurements,images and calculations discussed above may be performed before asubject undergoes a carotid endarterectomy (CEA), which is a surgicalprocedure used to reduce the likelihood, or possibly prevent stroke, bycorrecting stenosis (narrowing) in the common carotid artery.Endarterectomy is the removal of material on the inside (end-) of anartery. A blood vessel 4501 including a narrowing 4597 is illustrated,for example, in FIG. 45. A velocity field/profile may be calculated atvarious point in the blood vessel as illustrated in FIG. 46 before andafter the carotid endarterectomy to correct the narrowing 4597. Thus, bycomparing the measurements/quantities before and after the procedure,the success of the procedure may be determined. FIG. 47 is a graphillustrating fluid rate estimate along the diameter of the blood vessel4501 illustrated in FIGS. 45 and 46.

Although uses of the methods and systems discussed herein are discussedspecifically with respect to carotid endarterectomies, it will beunderstood that embodiments of the present inventive concept are notlimited to this configuration. For example, embodiments of the presentinventive concept may be used for the brain, colon, or any otherapplicable part of the subject that may benefit from the techniquesdiscussed herein.

Operations for a non-invasive method for measuring blood flow inprincipal vessels of a subject in accordance with some embodiments willbe discussed with respect to FIG. 48. Operations begin at block 4816 byilluminating a region of interest with a coherent light source, whereinthe coherent light source has a wavelength of from about 600 nm to about1100 nm. At least two speckle images of the region of interest aresequentially acquired during a fixed time period (block 4826). Temporaland spatial variation of pixel intensities of the at least two acquiredspeckle images are electrically evaluated to determine spatialdistribution of blood flow speed in the principal vessels and perfusiondistribution in tissue in the region of interest (block 4836).

A velocity field for the region of interest is calculated (block 4846).In some embodiments, the velocity field is calculated using equations(9) and (10) set out below. Blood flow speed in the region of interestbased on the calculated velocity field is calculated (block 4856). Thecalculated blood flow speed in the region of interest is compared to theblood flow speed determined using the acquired at least two speckleimages of the region of interest to verify results obtained using the atleast two speckle images (block 4866). Thus, embodiments of the presentinventive concept may be used to verify experimental results as will bediscussed further below.

It will be understood that the operations of blocks 4816, 4826, 4836,4846, 4856 and 4866 may be performed before and after a procedureperformed on the subject. The results before and after the procedure maybe compared to verify the success of the procedure in the subject.

In the cardiac space things happen so quickly that timing can benormalized, i.e. the velocity of flow is so fast, time may not be afactor. However, in other portions of the body, for example, theextremities things happen much slower (velocity of flow is much slower)and, therefore, timing is much more important. Thus, according toembodiments of the present inventive concept discussed herein, an EKGmay be used to synchronize data acquisition, which may allow relativecomparison of a same area over time. This synchronization may allow useof LSI methods discussed herein in many areas of the body beyond theheart. Use of the EKG as a synchronization tool may also enablegeneration of instantaneous and average measurements of flow and/orperfusion as will be discussed further herein with respect to FIGS. 49through 67.

As will be discussed, in accordance with some embodiments of the presentinventive concept experiments were performed on a human heart, bothstopped and beating. As will be discussed, speckle imaging techniquesdiscussed herein can not only be used for non-invasive determination ofblood flow speed in the principal vessels and to quantify perfusiondistribution in tissue in the region of interest from the LSCI image,but also to generate instantaneous and average flow and perfusion maps.Furthermore, as discussed above, EKG gating is used as a temporalstandard for analysis for acquisition of data both in the heart regionas well as other areas of the body where this information may be useful.Because the LSI data in accordance with embodiments discussed herein aresynchronized using EKG gating, LSI technology in accordance withembodiments of the present inventive concept will be discussed herein as“triggered LSI.” In other words, the data collection is synchronizedusing a metric, in embodiments discussed herein, the metric may be apatient's EKG and, thus, the acquisition of the LSI image is“triggered.”

Referring first to FIG. 49, experiments discussed herein were performedusing a SPY® Imaging System illustrated in FIG. 49. The SPY imagingsystem is offered by Novadaq Technologies Inc. The SPY system is easilymobile from room to room. Set up can be managed by the operating roomstaff. The articulating arm and camera head are controlled by thesurgeon. Image capture may accomplish in less than two minutes andimages can be replayed immediately for review.

Experiments in accordance with embodiments of the present inventiveconcept were performed using the SPY system as the SPY system was areadily available fluorescence imaging system. However, modificationswere made to the system for embodiments discussed herein. For example,no ICG dye was injected into the subject as the triggered LSI procedurediscussed herein does not use dye injection. Using the SPY systempresented various disadvantages, for example, the frame rate of the SPYdevice is slow, about 30 frames per second (fps), which the limits theamount of data that can be acquired. Furthermore, the wavelength of thelaser in the SPY device is about 810 nm, but fluorescence is about 830nm. There is a long pass filter in front of the camera that removes mostof the 810 nm light. Triggered LSI only uses 810 nm and the reflectedlight is decreased to 1/10,000 because of the filter. Thus, results maybe greatly improved with equipment optimized for triggered LSI. Asdiscussed, in the later examples and figures, the SPY system primarilyfunctioned in these experiments simply as the laser illumination source.As will be illustrated below, however, results obtained using the SPYdevice are capable of illustrating various aspects of the presentinventive concept as will be discussed below.

Although many the following figures are discussed with respect to astopped and/or beating heart, embodiments of the present inventiveconcept are not limited to this configuration. For example, embodimentsof the present inventive concept may be used in coronary artery bypass,cardiovascular, plastic, reconstructive, micro, organ transplant andgastrointestinal surgery as well as many other procedures that couldbenefit from the information provided by the inventive concept withoutdeparting from the scope of the present inventive concept.

Referring now to FIGS. 50A and 50B, the images illustrated therein wereobtained using LSI imaging using the modified SPY device on amotionless, non-beating heart using a heart-lung machine and no dye wasinjected into the subject. In particular, FIG. 50A illustrates a singleframe of LSI data and FIG. 50B illustrates an inverse laser speckletemporal contrast imaging (LSTCI) image in accordance with embodimentsof the present inventive concept. As illustrated in FIG. 50B, the leftanterior descending coronary artery (LAD) is clearly visible in theLSTCI image. The stopped heart (AKA antegrade cardioplegia) imaged inFIGS. 50A and 50B was with the patient on the cardiopulmonary bypasspump. The LAD in FIG. 50B has a high velocity flow. As discussed above,these images were obtained using the SPY device where reflective lightis decreased to 1/10,000 due to the filter. However, the LAD is stillvery visible in the LSTCI image of FIG. 50B. Although FIG. 50Billustrates a temporal contrast image, it will be understood that bothspatial and temporal contrast analyses are used in accordance withembodiments discussed herein.

Referring now to FIGS. 51A and 51B, the images illustrated therein wereobtained using LSI imaging using the modified SPY device on the samemotionless, non-beating heart as illustrated in FIGS. 50A and 50B, butfluid was injected retrograde (AKA retrograde cardioplegia) for theseimages. In particular, FIG. 51A illustrates a single frame of LSI dataand FIG. 51B illustrates an LSTCI image in accordance with embodimentsof the present inventive concept. As illustrated in FIG. 51B, the leftanterior descending coronary artery (LAD) is clearly visible in theLSTCI image and has lower velocity of flow than the LAD illustrated inFIG. 50B. The stopped heart (AKA retrograde cardioplegia) imaged inFIGS. 51A and 51B was with the patient on the cardiopulmonary bypasspump. Thus, embodiments of the present inventive concept can be used todetect differences in velocity at different points in time in the samevessels, for example, the LAD illustrated in FIGS. 50B (antegrade flow,higher velocity) and 51B (retrograde flow, lower velocity).

Referring now to FIGS. 52A and 52B, a conventional near-infraredfluorescence SPY image is compared to a LSTCI image in accordance withembodiments discussed herein. In particular, FIG. 52A illustrates oneframe of a conventional SPY image system in venous phase. As illustratedin FIG. 52A, the bypass graft to the LAD is identified in the SPY image.FIG. 52B is an image generated using inverse LSTCI in accordance withembodiments discussed herein. The LAD is pointed out in FIG. 52B. Whenthese images are compared, it is clear that the image generated usingembodiments of the present inventive concept of FIG. 52B is very similarto the SPY image of FIG. 52A. In other words, embodiments of the presentinventive concept are illustrating the imaged anatomy accurately.Furthermore, as discussed above, using triggered LSI technology inaccordance with embodiments discussed herein, different velocities inthe arteries and veins can be illustrated. SPY systems only provideimages of flow (1) or no flow (0), but cannot depict differentvelocities of flow as discussed herein.

Information related to the velocity of flow may be very useful to, forexample, a surgeon placing a graft on the heart. In real time,embodiments of the present inventive concept may allow the surgeon toaccess the velocity of flow in the arteries and veins around the graftbefore the placement of the graft and after placement of the graft,thus, allowing an assessment of the surgeries success in real time. Thisinformation may be very valuable as will be discussed further below.

As discussed above, results in accordance with various embodimentsdiscussed herein are affected by the quality of the equipment used toobtain the various measurements and images. Referring now to FIGS. 53Athrough 53D, in some embodiments images were obtained using a cameraoperating at 60 fps, twice that of the original camera used (30 fps).FIG. 53A is an illustration of the SPY device including the new cameraconfigured to capture images at 60 fps. FIG. 53B is a larger view of thecamera 5330 attached to the arm of the SPY device. FIG. 53C illustratesthe system covered in plastic to provide a sanitary environment in theoperating room. FIG. 53D illustrates the field of view (FOV) captured bythe camera, which in this particular example is the heart of thepatient.

Referring now to FIGS. 54A through 54C, images obtained from anon-beating heart using the system with the 60 fps camera discussedabove will be discussed. In particular, FIG. 54A is a single imageframe, FIG. 54B is an inverse LSSCI image obtained from images capturedusing the 60 fps camera and FIG. 54C is an inverse LSTCI image obtainedfrom images captured using the 60 fps camera. Comparing imagesillustrated in FIG. 54A-54C and the images of FIGS. 50A-52B, it is clearthat the structures of the heart are more clearly shown in the imagesobtained using the camera with higher fps. It is also clear that theLSTCI image is typically a better image than the LSSCI image.

Referring now to FIGS. 55A through 55D, images obtained in a beatingheart using the camera that obtains images at 60 fps camera will bediscussed. FIG. 55A is a single image frame, FIG. 55B is an inverseLSSCI image obtained from images captured using a 60 fps camera and FIG.55C is an inverse LSTCI image obtained using images generated with the60 fps camera. Thus, embodiments of the present inventive concept may beused to capture images in a moving heart, thus, enabling instantaneousflow and perfusion measurements of the various arteries, veins, andtissues. However, as is clear from the images of FIGS. 55B and 55C, acamera that captures images at a higher fps rate would be desirable. Forexample, a camera speed of 600 fps would result in far more detailedimages. Furthermore, many of the black areas of the images may be causedby inadvertent reflection off the sterile plastic cover illustrated inFIG. 53C that is provided for a sterile operating environment.

Referring now to FIG. 55D, a graph of average intensity vs. time inseconds will be discussed. This graph tracks the movement of the surfaceof the heart during the individual heartbeats, and acts as a surrogatefor the EKG. The object distance between the camera lens and the heartsurface changes with the heart cardiac cycle, moving closer and fartheraway within each cycle, causing the average intensity of an individualframe to fluctuate in this cyclical pattern shown in FIG. 55D. Inaddition, the average intensity vs. time curve can be filtered to be asurrogate EKG signal in accordance with embodiments discussed herein. Inparticular, the graph illustrates instantaneous LSI velocitymeasurements at any point in time in the cardiac cycle, for example, asindicated by the sequential squares on the peak at 2 secs. However, itis unclear which portions of the graph correspond to systole, i.e.,indicating the maximum arterial pressure occurring during contraction ofthe left ventricle of the heart, and diastole, i.e. indicating thelowest arterial pressure during the interval between heartbeats. Todetermine which portions of the graph correspond to which cycles of theheart a constant is needed, for example, an EKG gating can be used as atemporal standard for analysis.

By observing the actual beating of the heart over a period of time, andincorporating the EKG gating concepts, it was determined which portionsof the graph correspond to the relevant time-points in the cardiaccycles of the heart as illustrated in FIGS. 56A and 56B. FIG. 56A is astill shot from a movie of a moving heart used to obtain the graph ofaverage intensity vs. time in seconds illustrating instantaneous LSIvelocity measurements of FIG. 56B. As illustrated in FIG. 56B, bywatching the moving heart and using EKG gating as discussed herein, theportions of the graph corresponding to diastole and systole wereidentified. The EKG gating concepts were used to separate the diastolicand systolic phases.

In embodiments discussed above, the EKG may be used to find, forexample, a specific phase in the cardiac cycle where heart movement isminimized (diastole) and an instantaneous flow-perfusion map can begenerated during this very short time period. In accordance withembodiments discussed herein, instantaneous flow and perfusion maps canbe generated at any time in the cardiac cycle using EKG gating asdiscussed herein. In particular, embodiments of the present inventiveconcept take advantage of the fact that every patient in an operatingroom is connected to an EKG machine, and EKG machines are readilyavailable in Clinics and other outpatient settings. Thus, the EKG can beused to synchronize the data collection and the analysis of the imagingdata.

Average flow and perfusion maps can also be generated over severalcardiac cycles used on the EKG gating embodiments discussed herein.Although embodiments discussed above use EKG synchronization in cardiacapplications, embodiments of the present inventive concept are notlimited to this configuration. EKG synchronization can be used as amethodology for non-cardiac triggered LSI applications to compare flowand perfusion using instantaneous/average flow and perfusion maps inother parts and organs of the body, i.e., non-cardiac portions.

In particular, when dealing with the heart (cardiac conditions andprocedures) things happen pretty quickly, and an event can occur and beover in milliseconds. Also, blood flow varies dramatically (from zero tomaximal). Thus, in cardiac applications, both instantaneous flow andaverage flow determinations typically require a method for locating andreferencing time vs. what is happening physiologically. In other partsof the body outside the heart, blood flow is more constant, butinstantaneous flow and average flow determinations still need a methodfor locating and referencing time vs. what is happening physiologically,i.e., some physiologic timer. Thus, in accordance with embodiments ofthe present inventive concept, the EKG can be used as a trigger for datacollection, acquisition and analysis. Because things happen much sloweroutside of the heart, this EKG synchronization is also necessary as thisphysiologic timer for accurate results and can be used as a third railfor data collection.

Real time data collection and creation of instantaneous flow andperfusion maps may save a lot of time in a research environment. Forexample, if the researcher is studying the effect ofvasoconstrictive/vasodialative drugs on the human body, i.e. how fastdoes the drug take effect, how long does it last and the like. TriggeredLSI technology can be used on the area of interest in the body and flowand perfusion maps may be generated before, during and afteradministration of the drugs to access the necessary aspects. This sametype of analysis may take days in the lab with multiple blood draws andscans being necessary.

Referring now to FIG. 57, an example of how EKG gating could generate aninstantaneous flow and perfusion map is illustrated. The EKG tracingshown is used to identify any selected component of the cardiac cycle,for example systole (Panel A) or diastole (Panel B). Because of this,the EKG can be used to trigger data acquisition, check for timing withinthe cycle, and used as an external temporal reference in non-cardiacapplications of the embodiments discussed herein. As discussed above, aninstantaneous flow and perfusion map may be generated at any time in acardiac cycle using EKG synchronization techniques in accordance withembodiments discussed herein.

Referring now to FIGS. 58A through 59D, instantaneous flow measurementsat times approximating end-diastole and end-systole will be discussed.The images in FIGS. 58A-C and 59A-C were obtained in a beating heartusing the camera that obtains images at 60 fps camera. FIGS. 58A and 59Aare single image frames, FIGS. 58B and 59B are inverse LSSCI imagesobtained from images generated using the 60 fps camera and FIGS. 58C and59C are inverse LSTCI images obtained using images generated with the 60fps camera. Thus, as discussed above, embodiments of the presentinventive concept may be used to capture images in a moving heart,allowing instantaneous flow measurements from the various arteries,veins and tissues. FIGS. 58A-58C are images representing an anteriorwall of the heart at a time approximating end-diastole (max blood flow).FIG. 58C specifically illustrates the graft inserted by the surgeon intothe LAD. FIG. 58D is a graph of average intensity vs. time in secondsand illustrates instantaneous LSI velocity measurements at any point intime in the cardiac cycle. The highlighted portions of the graphcorrespond to velocities at a time approximating end-diastole. Thisgraph is generated in a similar fashion to that described in FIG. 55D.FIGS. 59A through 59C are images representing an anterior wall of theheart at a time approximating end-systole (min blood flow). FIG. 59Cspecifically illustrates the graft inserted by the surgeon and the LAD.FIG. 59D is a graph of average intensity vs. time in seconds andillustrates instantaneous LSI velocity measurements at any point in timein the cardiac cycle. The highlighted portions (squares) of the graphcorrespond to velocities at a time approximating end-systole. This graphis generated in a similar fashion to that described in FIG. 55D. As isclear from comparing these graphs, differences in velocities can bemeasured in a beating heart using the LSI method discussed herein. Thedifferences in blood flow can even be seen by comparing the LAD of FIG.58C and the LAD of FIG. 59C.

Referring now to FIG. 60, this is an example of how average flow andperfusion can be generated from the EKG triggering and analysis. Asdiscussed above, an average flow and perfusion map may be generated bycombining image data from two or more several cardiac cycles using EKGsynchronization techniques in accordance with embodiments discussedherein. This EKG trace illustrates capture of data during systolesequentially from three cardiac cycles.

Referring now to FIGS. 61A through 62D, average flow measurements indiastole and systole during multiple cardiac cycles will be discussed.The images in FIGS. 61A-C and 62A-C were obtained in a beating heartusing the camera that obtains images at 60 fps camera. FIGS. 61A and 62Aare single image frames, FIGS. 61B and 62B are inverse LSSCI imagesobtained using the 60 fps camera and FIGS. 61C and 62C are inverse LSTCIimages obtained using images generated with the 60 fps camera. Thus, asdiscussed above, embodiments of the present inventive concept may beused to capture images in a moving heart, allowing average flowmeasurements of the various arteries, veins and tissues during multiplecardiac cycles to be generated. FIGS. 61A-61C are images representing ananterior wall of the heart at a time approximating end-diastole (maxblood flow) during multiple cardiac cycles. FIG. 61D is a graph ofaverage intensity vs. time in seconds and illustrates average LSIvelocity measurements during multiple cardiac cycles. This graph isgenerated in identical fashion to FIG. 55D, but in this case illustratesimage acquisition data at a time approximating end-diastole from 9sequential cardiac cycles for analysis. The highlighted portions of thegraph correspond to velocities at a times approximating end-diastolefrom the multiple cycles. FIGS. 62A-62C are images representing ananterior wall of the heart at a time approximating end-systole (minblood flow). FIG. 62D is a graph of average intensity vs. time inseconds and illustrates average LSI velocity measurements at any pointin time in the cardiac cycle. This graph was generated in a similarfashion to FIG. 55D, but in this case illustrates image acquisition dataat a time approximating end-systole from 8 sequential cardiac cycles foranalysis. The highlighted portions of the graph correspond to velocitiesat a time approaching end-systole. As is clear from comparing thesegraphs, differences in velocities can be measured in a beating heartusing the LSI method discussed herein. To compare flow and perfusionmaps, for example, before and after an intervention, EKG synchronizationis an ideal technique. As discussed above, every patient in an operatingroom has an EKG. In addition, EKG machines are readily available in allclinic and outpatient environments. An EKG is a standardized,physiologic timing template that can be used as a baseline forcomparison. This is a positive because in physiology, time phase is notstandard across LSI clinical applications. EKG also creates theanalytical basis for comparison of instantaneous flow and perfusionmaps, because the flow and perfusion patterns will vary based on thephysiology/pathophysiology of blood flow and perfusion. EKGsynchronization is an ideal way to link the flow and perfusion to anindependent, objective benchmark, i.e, a specific cardiac phase.

EKG also creates an analytical basis for comparison of average flow andperfusion maps because the objective precision of the EKG is ideal fordefining the starting and ending points of the averaging process versussimply finding a random starting point and averaging a few seconds ofimaging data.

Referring now to FIGS. 63A through 63D, embodiments of the presentinventive concept using EKG gating concepts to provide informationrelated to graft procedures will be discussed. The images in FIGS. 63Athrough 63C were obtained in a beating heart using the camera thatobtains images at 60 fps camera. FIG. 63A is a single image frame, FIG.63B is an inverse LSSCI image obtained using images generated by the 60fps camera and FIG. 63C is an inverse LSTCI image obtained using imagesgenerated with the 60 fps camera. In embodiments illustrated in FIGS.63A-C, a clamp has been placed on the bypass graft to simulate nativeblood flow through the arteries, veins and tissues before the bypassgraft was inserted into the patient. FIG. 63D is a graph of averageintensity vs. time in seconds and illustrates evolution of perfusionchange induced by bypass grafting. This graph was generated in a similarfashion to FIG. 55D, with three cycles analyzed at a time approximatingend-diastole (maximal coronary blood flow). As illustrated in FIG. 63C,there is no flow through the graft when the clamp on the graft and thus,the flow through the LAD would be equivalent to the flow before thegraft was inserted into the patient.

In stark contrast, FIGS. 64A through 64D, illustrate embodiments wherethe clamp has been removed from the graft and blood was allowed to flowthrough the graft. The images in FIGS. 64A through 64C were obtained ina beating heart using the camera that obtains images at 60 fps camera.FIG. 64A is a single image frame, FIG. 64B is an inverse LSSCI imageobtained using imaged generated by the 60 fps camera and FIG. 64C is aninverse LSTCI image obtained using images generated with the 60 fpscamera. As illustrated in FIG. 64C, there is blood flow through thegraft and blood flow through the LAD has increased. FIG. 64D is a graphof average intensity vs. time in seconds and illustrates evolution ofperfusion change induced by bypass grafting. This graph was generated ina similar fashion to FIG. 55D, with three cycles again analyzed at atime approximating end-diastole. Thus, with this EKG-concept timing,FIG. 64C can be directly compared to FIG. 63C, with the imagingdifference being the flow and velocity resulting from the bypass graftto the heart vessel.

This comparison between FIGS. 63C and 64C illustrates the concept of EKGtriggering for data acquisition, and EKG timing for average velocitiesand comparative analysis of those average velocities. Comparison ofinstantaneous velocities could be accomplished in the same manner, inany physiologic circumstance where study of blood flow and tissueperfusion was indicated.

However, as discussed above, embodiments of the present inventiveconcept do not have to be used in cardiac applications. In other words,LSI technology can be extended to other applications. Physiologically,the cardiac application is the most complicated for flow and perfusionanalysis. Other applications such as wound healing, tissuereconstruction, vascular surgery and transplantation have fewerchallenges for triggered LSI imaging.

Referring now to FIGS. 65A through 65C, images of a hand of a persongenerated using embodiments discussed herein will be discussed. FIG. 65Ais a single image frame, FIG. 65B is an inverse LSSCI image obtainedusing images generated by the 60 fps camera and FIG. 65C is an invertedLSTCI obtained using images generated with the 60 fps camera. Theseimages clearly illustrate microvascular blood flow in the human handusing LSI technology in accordance with embodiments discussed herein.

Using triggered LSI in accordance with embodiments of the presentinventive concept, real time flow and perfusion maps may be generatedduring procedures. Thus, a success of a procedure may be immediatelyassessed and remedied, if necessary. Flow is measured as ccs/minute.Since the frame rate of a camera is slower than a processor, forexample, data processing system 200 of FIG. 2A, in a communicationsdevice, the algorithm for generating the flow and perfusion diagrams canbe hard coded into the system to provide real time information to theperson performing the procedure. As discussed above, this real timeinformation can be invaluable.

Referring now to FIGS. 66 and 67, flowcharts illustrating operations inaccordance with various embodiments of the present inventive conceptwill be discussed. Referring first to FIG. 66, operations begin at block6615 illuminating a region of interest of a subject with a coherentlight source. At least two speckle images are acquired of the region ofinterest (block 6625). The at least two speckle images are acquired insynchronization along with a simultaneously acquired EKG tracing as thetiming signal. The at least two acquired speckle images areelectronically processed based on the temporal variation of the pixelintensities in the at least two acquired speckle images to generate alaser speckle contrast imaging (LSCI) image, determine distribution ofblood flow speed in principal vessels and quantify perfusiondistribution in tissue in the region of interest from the LSCI image(block 6635). As discussed above, in accordance with embodimentsdiscussed herein, the LSCI image enables detection of different bloodflow speeds, which is not possible in conventional methods.

Referring now to FIG. 67, operations begin at block 6715 by illuminatinga region of interest of a subject with a coherent light source. At leasttwo speckle images are acquired of the region of interest (block 6725).The at least two speckle images are acquired in synchronization, alongwith a simultaneously acquired EKG trace as the timing signal (block6725). Using EKG-based timing to select frames for instantaneous oraverage flow analysis, the at least two acquired speckle images areelectronically processed based on the temporal, spatial, or combinationof both contrast variation of the pixel intensities to generate a laserspeckle contrast imaging (LSCI) image (block 6735). This LSCI image isproportional to the flow speed of the region of interest. Using EKG asthe corresponding physiologic timing locater LSCI is used to determinethe instantaneous flow speed in the principal vessels and/or perfusionin the tissue at any time in one EKG cycle and average flow speed in theprincipal vessels and/or perfusion in the tissue in two or more EKGcycles (block 6735). The flow speed image is further input into ananalytical model such as fluid dynamic model to generate manymeasurements such as volume flow rate in the principal vessels andquantified perfusion map in the tissue, flow direction and pressuredistribution etc. (block 6745). The instantaneous flow rate and/orperfusion maps or the average flow rate and/or perfusion maps generatedat pre and post operation are compared to determine efficacy of atreatment (6755).

It will be understood that the region of interest may be a cardiacregion of interest or a non-cardiac region of interest without departingfrom the scope of the present inventive concept.

As discussed above, embodiments of the present inventive concept may beused to not only determine blood flow velocity, but blood flow rates andvelocity integrated across cross section areas of the blood vessels. Themeasurements are obtained according to heart beat timing signals of anelectrocardiogram (EKG). The results of the measurement of blood flowdistribution are validated using triggered LSI and enhanced byhydrodynamic modeling of blood flows.

The foregoing is illustrative of the present inventive concept and isnot to be construed as limiting thereof. Although a few exemplaryembodiments of the present inventive concept have been described, thoseskilled in the art will readily appreciate that many modifications arepossible in the exemplary embodiments without materially departing fromthe novel teachings and advantages of this inventive concept.Accordingly, all such modifications are intended to be included withinthe scope of the inventive concept as defined in the claims. In theclaims, means-plus-function clauses, where used, are intended to coverthe structures described herein as performing the recited function andnot only structural equivalents but also equivalent structures.Therefore, it is to be understood that the foregoing is illustrative ofthe present inventive concept and is not to be construed as limited tothe specific embodiments disclosed, and that modifications to thedisclosed embodiments, as well as other embodiments, are intended to beincluded within the scope of the appended claims. The inventive conceptis defined by the following claims, with equivalents of the claims to beincluded therein.

That which is claimed is:
 1. A non-invasive method for determining bloodflow distribution in a region of interest, the method comprising:illuminating a region of interest of a subject with a coherent lightsource; sequentially acquiring at least two speckle images of the regionof interest, wherein sequentially acquiring the at least two speckleimages comprises acquiring the at least two speckle images insynchronization with motion of the heart of the subject; andelectronically processing the at least two acquired speckle images basedon the temporal variation of the pixel intensities in the at least twoacquired speckle images to generate a laser speckle contrast imaging(LSCI) image, determine distribution of blood flow speed in principalvessels and quantify perfusion distribution in tissue in the region ofinterest from the LSCI image, wherein the LSCI image enables detectionof different blood flow speeds.
 2. The method of claim 1, whereinsequentially acquiring the at least two speckle images comprisesacquiring the at least two speckle images in synchronization with motionof the heart of the subject further comprises: electronically monitoringan EKG cardiac cycle of the subject; and electronically synchronizingacquisition of speckle images with the EKG signals.
 3. The method ofclaim 2, wherein the region of interest is a beating heart, the methodfurther comprising generating instantaneous flow and/or perfusion mapsfor the region of interest at any time during a cardiac cycle using theEKG signals to synchronize data acquisition.
 4. The method of claim 3,further comprising comparing instantaneous flow and/or perfusion mapsgenerated at first and second times to determine efficacy of atreatment.
 5. The method of claim 4, wherein the first time is a timebefore a treatment is administered and wherein the second time is a timeafter the treatment is administered.
 6. The method of claim 2, whereinthe region of interest is a beating heart, the method further comprisinggenerating average flow and/or perfusion maps for the region of interestover two or more cardiac cycles using EKG signals to synchronize dataacquisition.
 7. The method of claim 6, further comprising comparingaverage flow and/or perfusion maps generated at first and second timesto determine efficacy of a treatment.
 8. The method of claim 7, whereinthe first time is a time before a treatment is administered and whereinthe second time is a time after the treatment is administered.
 9. Themethod of claim 2, wherein the region of interest is a non-cardiacregion of the subject, the method further comprising generatinginstantaneous flow and/or perfusion maps at any time during dataacquisition using the EKG signals to synchronize data acquisition. 10.The method of claim 9, further comprising comparing instantaneous flowand/or perfusion maps generated at first and second times to determineefficacy of a treatment.
 11. The method of claim 10, wherein the firsttime is a time before a treatment is administered and wherein the secondtime is a time after the treatment is administered.
 12. The method ofclaim 2, wherein the region of interest is a non-cardiac region of thesubject, the method further comprising generating average flow and/orperfusion maps over two or more periods of data acquisition using EKGsignals to synchronize data acquisition.
 13. The method of claim 12,further comprising comparing average flow and/or perfusion mapsgenerated at first and second times to determine efficacy of atreatment.
 14. The method of claim 13, wherein the first time is a timebefore a treatment is administered and wherein the second time is a timeafter the treatment is administered.
 15. The method of claim 1, whereinthe coherent light source has a wavelength of from about 600 nm to about1100 nm.
 16. A non-invasive method for determining blood flowdistribution in a region of interest, the method comprising:illuminating a region of interest of a subject with a coherent lightsource; sequentially acquiring at least two speckle images of the regionof interest, wherein sequentially acquiring the at least two speckleimages comprises acquiring the at least two speckle images insynchronization with motion of the heart of the subject; electronicallyprocessing the at least two acquired speckle images based on thetemporal variation of the pixel intensities in the at least two acquiredspeckle images to generate a laser speckle contrast imaging (LSCI)image, determine distribution of blood flow speed in principal vesselsand quantify perfusion distribution in tissue in the region of interestfrom the LSCI image; generating one of instantaneous flow and/orperfusion maps for the region of interest at any time during dataacquisition using EKG signals to synchronize data acquisition andaverage flow and/or perfusion maps over two or more periods of dataacquisition using the EKG signals to synchronize data acquisition; andcomparing one of the instantaneous flow and/or perfusion maps and theaverage flow and/or perfusion maps generated at first and second timesto determine efficacy of a treatment.
 17. The method of claim 16,wherein the region of interest is a beating heart.
 18. A non-invasivesystem for determining blood flow distribution in a region of interest,the system comprising: a coherent light source configured to illuminatea region of interest of a subject; a camera in communication with thecoherent light source that is configured to sequentially acquire atleast two speckle images of the region of interest, wherein acquisitionof the at least two speckle images is synchronized with motion of theheart of the subject; and a data processing circuit configured toevaluate the temporal variation of the pixel intensities in the at leasttwo acquired speckle images to generate a laser speckle contrast imaging(LSCI) image, determine distribution of blood flow speed in theprincipal vessels and quantify perfusion distribution in tissue in theregion of interest in the heart from the LSCI image, wherein the LSCIimage enables detection of different blood flow speeds.
 19. The systemof claim 18, wherein the data processing circuit is further configuredto: electronically monitor an EKG cardiac cycle of the subject; andelectronically synchronize acquisition of speckle images with the EKGsignals.
 20. The system of claim 19, wherein the region of interest is abeating heart, the system further comprising a modeling moduleconfigured to generate instantaneous flow and/or perfusion maps for theregion of interest at any time during a cardiac cycle using the EKGsignals to synchronize data acquisition.
 21. The system of claim 20,wherein the data processing circuit is further is configured to compareinstantaneous flow and/or perfusion maps generated at first and secondtimes to determine efficacy of a treatment.
 22. The system of claim 19,wherein the region of interest is a beating heart, the system furthercomprising a modeling module configured to generate average flow and/orperfusion maps for the region of interest over two or more cardiaccycles using EKG signals to synchronize data acquisition.
 23. The systemof claim 22, wherein the data processing circuit is further configuredto compare average flow and/or perfusion maps generated at first andsecond times to determine efficacy of a treatment.
 24. The system ofclaim 19, wherein the region of interest is a non-cardiac region of thesubject, the system further comprising a modeling module configured togenerate instantaneous flow and/or perfusion maps at any time duringdata acquisition using the EKG signals to synchronize data acquisition.25. The system of claim 24, wherein the data processing circuit isfurther configured to compare instantaneous flow and/or perfusion mapsgenerated at first and second times to determine efficacy of atreatment.
 26. The system of claim 19, wherein the region of interest isa non-cardiac region of the subject, the system further comprising amodeling module configured to generate average flow and/or perfusionmaps over two or more periods of data acquisition using EKG signals tosynchronize data acquisition.
 27. The method of claim 26, wherein thedata processing circuit is further configured to compare average flowand/or perfusion maps generated at first and second times to determineefficacy of a treatment.
 28. A computer program product for determiningblood flow distribution in a region of interest, the computer programproduct comprising: a non-transitory computer-readable storage mediumhaving computer-readable program code embodied in the medium, thecomputer-readable program code comprising: computer readable programcode configured to illuminate a region of interest of a subject with acoherent light source; computer readable program code configured tosequentially acquire at least two speckle images of the region ofinterest, wherein sequentially acquiring the at least two speckle imagescomprises acquiring the at least two speckle images in synchronizationwith motion of the heart of the subject; computer readable program codeconfigured to electronically process the at least two acquired speckleimages based on the temporal variation of the pixel intensities in theat least two acquired speckle images to generate a laser specklecontrast imaging (LSCI) image, determine distribution of blood flowspeed in principal vessels and quantify perfusion distribution in tissuein the region of interest from the LSCI image; computer readable programcode configured to generate one of instantaneous flow and/or perfusionmaps for the region of interest at any time during data acquisitionusing EKG signals to synchronize data acquisition and average flowand/or perfusion maps over two or more periods of data acquisition usingthe EKG signals to synchronize data acquisition; and computer readableprogram code configured to compare one of the instantaneous flow and/orperfusion maps and the average flow and/or perfusion maps generated atfirst and second times to determine efficacy of a treatment.
 29. Anon-invasive method for determining blood flow distribution in a regionof interest, the method comprising: illuminating a region of interest ofa subject with a coherent light source; sequentially acquiring at leasttwo speckle images of the region of interest, wherein sequentiallyacquiring the at least two speckle images comprises acquiring the atleast two speckle images in synchronization in synchronization with anelectrocardiogram (EKG); selecting at least two speckle images usingEKG-based timing; processing the selected images to determine aninstantaneous/average flow speed (centimeter/second) in the region ofinterest using at least one of a temporal contrast algorithm and aspatial contrast algorithm, wherein the EKG is used at any time duringdata acquisition and analysis to select frames to be processed, tolocate the instantaneous flow speed in one or more EKG cycles, and/or totarget the beginning and ending time of average flow speed analyses;inputting the instantaneous/average flow speed image into the analysismodel to generate a flow rate (cubic centimeter/second) map in theprincipal vessels and a perfusion map in the microvascular structure;and generating a direction of flow and pressure.